SAFETY AND PHARMACOKINETICS OF PRAZIQUANTEL IN EUROPEAN POND TURTLES (EMYS ORBICULARIS).

Spirorchiidosis, caused by blood flukes of the genus Spirorchis, is a disease of great concern for the critically endangered European pond turtle (EPT; Emys orbicularis) in Switzerland. The endogenous life cycle of the parasite often leads to systemic inflammatory reactions, thrombosis, and death. Praziquantel (PZQ) is the treatment of choice against adult Spirorchis spp. in green (Chelonia mydas) and in loggerhead (Caretta caretta) sea turtles and is therefore considered for the treatment of EPT. This study aimed to establish a safe, easily applicable PZQ treatment for EPT, based on pharmacokinetics and tolerability. Three application methods were tested in a total of 12 adult EPT. Each turtle received a total of 75 mg/kg PZQ (three doses of 25 mg/kg in 3-h intervals [q3h × 3]) via IM (n = 3 turtles), SC (n = 3 turtles), or PO (n = 6 turtles) administration. Blood was collected 3, 6, 24, and 48 h after the first administration to determine the plasma concentration of PZQ using high-performance liquid chromatography coupled to mass spectrometry. Maximum measured R-PZQ concentrations (Cmax) were reached after 6 h. The mean Cmax of the total PZQ (sum of R- and S-PZQ) in the PO-treated EPT group was 1,929 ng/ml. Significantly higher concentrations were measured after IM and SC injection (mean Cmax of total PZQ = 12,715 ng/ml and 10,114 ng/ml, respectively). Transient side effects were evident after IM administration (local swelling and lameness), whereas no adverse drug effects were observed after PO and SC administration. Based on these results and the ease of administration to EPT, SC injection of PZQ at 25 mg/kg q3h times 3 serves as promising treatment application for the future.

What is the role of Swiss domestic cats in environmental contamination with Echinococcus multilocularis eggs?

BACKGROUND: The role of the domestic cat as definitive host for Echinococcus multilocularis and thus in environmental contamination with eggs has not yet been entirely resolved. This study aimed to assess the prevalence of E. multilocularis and other gastrointestinal parasites in Swiss domestic cats and to compare the diagnostic sensitivity of different methods for the detection of intestinal taeniid infection. METHODS: Faecal samples from 146 cats were included in the study. Faecal samples only were available from 55 cats; for the other 91 cats, necropsy was performed in addition to faecal sample testing. All (n = 146) faecal samples were analysed by a combined sedimentation/flotation technique (44% ZnCl2) and by the sodium acetate-acetic acid-formalin (SAF) sedimentation technique; when sufficient material was available (n = 121 samples) the Baermann-Wetzel technique was also used. Additionally, all samples were analysed by two coproantigen (copro)-quantitative PCRs (qPCR): (i) a multiplex qPCR able to detect and differentiate between E. multilocularis, Echinococcus granulosus sensu lato and Taenia spp./other cestodes (CEST-qPCR) and (ii) an E. multilocularis-specific qPCR (EM-qPCR). Finally, the intestines were examined macroscopically and microscopically for parasite stages at necropsy (n = 91) and using an intestinal scraping technique (IST) (n = 64). RESULTS: Of the 146 cats examined, 24 (17.1%) were infected by intestinal parasites, namely Hydatigera (syn. Taenia) taeniaeformis (8.9%), Toxocara cati (6.1%), Capillaria sp. (3.4%), hookworms (3.4%), Mesocestoides litteratus (1.4%), Giardia sp. (1.4%), Cystoisospora rivolta (1.4%), Cystoisospora felis (0.7%), Toxoplasma gondii (0.7%), Hammondia hammondi (0.7%) and Strongyloides sp. (0.7%). Necropsy and the IST revealed adult H. taeniaeformis in 12 animals, of which eight faecal samples were positive by the CEST-qPCR (sensitivity = 67%) and six samples by the sedimentation/flotation technique (sensitivity = 50%). No E. multilocularis infection was detected in the sampled cats. Using Bayesian latent class analysis, the mean posterior prevalence probability was 0.0% (95% confidence interval 0-0.83%) for E. multilocularis. CONCLUSIONS: There was no evidence of E. multilocularis infection among the 146 cats examined, suggesting that the prevalence of this parasite is low (< 1%) in the Swiss domestic cat population. Nonetheless, some of the sampled cats were infected by parasites that have rodents as intermediate hosts, demonstrating successful predation by these cats, and some were infected with zoonotic parasites. Cats therefore should not be disregarded as potential hosts for E. multilocularis and other zoonotic parasites.

Short communication: Efficacy of albendazole in Echinococcus multilocularis-infected mice depends on the functional immunity of the host.

Alveolar echinococcosis (AE) is a deadly parasitic disease that requires lifelong treatment with albendazole. Development of host immunity is pivotal with regard to the clinical outcome of AE, but its influence on conventional albendazole treatment is unknown. Using T-cell deficient athymic nude mice, we demonstrated that functional immunity is required for albendazole to be efficacious against murine AE. These results call for attention given the increasing number of immunocompromised patients with AE.

Foxp3+ T Regulatory Cells as a Potential Target for Immunotherapy against Primary Infection with Echinococcus multilocularis Eggs.

Alveolar echinococcosis (AE) is a lethal disease caused by infection with the metacestode stage of the helminth Echinococcus multilocularis, which develops into a tumorlike mass in susceptible intermediate hosts. The growth potential of this parasite stage is directly linked to the nature of the surrounding periparasitic immune-mediated processes. In a first step (experiment 1), mice were orally infected with E. multilocularis eggs, to be used for assessing the hepatic expression profiles of 15 selected cytokine and chemokine genes related to acquired immunity from 21 to 120 days postinfection. The early stage of infection in immunocompetent animals was marked by a mixed Th1/Th2 immune response, as characterized by the concomitant presence of gamma interferon (IFN-γ) and interleukin-4 (IL-4) and their related chemokines. At the late stage of AE, the profile extended to a combined tolerogenic mode including Foxp3, IL-10, and transforming growth factor beta (TGF-ß) as key components. In a second step (experiment 2), the effect of T regulatory cell (Treg) deficiency on metacestode growth was assessed in E. multilocularis-infected DEREG (depletion of regulatory T cells) mice upon induction of Treg deficiency with diphtheria toxin (DT). The parasite lesions were significantly smaller in the livers of treated mice than in corresponding control groups. Foxp3+ Tregs appear to be one of the key players in immune-regulatory processes favoring metacestode survival by affecting antigen presentation and suppressing Th1-type immune responses. For these reasons, we suggest that affecting Foxp3+ Tregs could offer an attractive target in the development of an immunotherapy against AE.

Serratospiculosis in Captive Peregrine Falcons (Falco peregrinus) in Switzerland.

Infection with Serratospiculum species was identified in a captive peregrine falcon (Falco peregrinus) in Switzerland. Pathologic and parasitologic examination results revealed generalized severe granulomatous airsacculitis, with intralesional adults, larvae, and eggs of Serratospiculum species. Subsequently, an individual coprological analysis of the remaining 15 falcons (peregrine falcons and gyrfalcons [Falco rusticolus]) from the same owner was performed. Eggs of Serratospiculum species (4 birds) and Capillaria species (11 birds), and oocysts of Caryospora species (1 bird) were detected. Treatment with ivermection (2 mg/kg SC) was effective, as none of the falcons excreted Serratospiculum species eggs 10 days after one dose. To our knowledge, this is the first report of infection with Serratospiculum species in captive falcons in Europe.

Besnoitia besnoiti lytic cycle in vitro and differences in invasion and intracellular proliferation among isolates.

BACKGROUND: Bovine besnoitiosis, caused by the protozoan Besnoitia besnoiti, reduces productivity and fertility of affected herds. Besnoitiosis continues to expand in Europe and no effective control tools are currently available. Experimental models are urgently needed. Herein, we describe for the first time the kinetics of standardised in vitro models for the B. besnoiti lytic cycle. This will aid to study the pathogenesis of the disease, in the screening for vaccine targets and drugs potentially useful for the treatment of besnoitiosis. METHODS: We compared invasion and proliferation of one B. tarandi (from Finland) and seven B. besnoiti isolates (Bb-Spain1, Bb-Spain2, Bb-Israel, Bb-Evora03, Bb-Ger1, Bb-France, Bb-Italy2) in MARC-145 cell culture. Host cell invasion was studied at 4, 6, 8 and 24 h post infection (hpi), and proliferation characteristics were compared at 24, 48, 72, 96, 120, and 144 hpi. RESULTS: In Besnoitia spp., the key parameters that determine the sequential adhesion-invasion, proliferation and egress steps are clearly distinct from those in the related apicomplexans Toxoplasma gondii and Neospora caninum. Besnoitia spp. host cell invasion is a rather slow process, since only 50 % of parasites were found intracellular after 3-6 h of exposure to host cells, and invasion still took place after 24 h. Invasion efficacy was significantly higher for Bb-France, Bb-Evora03 and Bb-Israel. In addition, the time span for endodyogeny to take place was as long as 18-35 h. Bb-Israel and B. tarandi isolates were most prolific, as determined by the tachyzoite yield at 72 hpi. The total tachyzoite yield could not be predicted neither by invasion-related parameters (velocity and half time invasion) nor by proliferation parameters (lag phase and doubling time (dT)). The lytic cycle of Besnoitia was asynchronous as evidenced by the presence of three different plaque-forming tachyzoite categories (lysis plaques, large and small parasitophorous vacuoles). CONCLUSIONS: This study provides first insights into the lytic cycle of B. besnoiti isolates and a standardised in vitro model that allows screening of drug candidates for the treatment of besnoitiosis.

Coprological analyses on apparently healthy Alpine ibex (Capra ibex ibex) from two Swiss colonies.

To provide baseline parasitological data for health surveillance in free-ranging Alpine ibex (Capra ibex ibex), we assessed the endoparasite population and level of parasitism in apparently healthy ibex. Faecal samples from 148 ibex were collected between 2006 and 2008 in two different Swiss ibex colonies. They were analysed by coprology, including combined sedimentation/flotation method, sedimentation method, Baermann funnel technique and Ziehl-Neelsen staining. Gastrointestinal parasites and lungworms were identified in 100% and 81.8% of the examined animals, respectively. Highest prevalences were recorded for gastrointestinal strongylids other than Nematodirus/Marshallagia spp. (100%), Eimeria spp. (100%), Muellerius spp. (79.8%) and Nematodirus/Marshallagia spp. (79.0%). We report for the first time Cryptosporidium sp. in free-ranging Alpine ibex and Cystocaulus spp. in free-ranging ibex from Switzerland. On average, ibex were infected with 3.9 different parasites taxa (range: 1-8). Parasite prevalence and diversity varied significantly between sexes, study sites and seasons. Parasite egg output was low in 95.7% and moderate in 5.3% of the samples. Overall, the results indicate that Alpine ibex are widely infected with endoparasites and suggest that multiple infections are very common in apparently healthy populations. Furthermore, our data underline the potential influence of factors such as sex, study site and season on parasitological findings.