Early stage Fasciola hepatica infection in growing cattle: impact on weight gain and liver weight determined in flukicide efficacy studies.

Although the negative impact of liver fluke (Fasciola hepatica) infection on production and health in cattle is generally accepted, results of individual research have been variable, ranging from important negative impacts on the animal to minimal or no impact. To add information on the impact of F. hepatica infection in growing cattle, weight gain and liver weight of young experimentally infected animals from seven controlled efficacy studies were analyzed. In each study, fluke naïve animals were inoculated with approximately 450 to 500 F. hepatica encysted metacercariae, blocked on body weight and randomly assigned into one untreated group (controls) and groups which were administered an experimental flukicide when the flukes were 4 weeks old (migrating) and sacrificed 8 weeks thereafter (12 weeks after inoculation). Data of groups which demonstrated >90% reduction of fluke counts following treatment and groups left untreated (total 103 and 47 animals, respectively) were compared. There was a significant (p < 0.0001) negative association between fluke count and weight gain while fluke count and liver weight and fluke count and relative liver weight were positively associated (p < 0.0001). Over the 8-week post-treatment period, flukicide-treated cattle had almost 15% more weight gain than the controls (50.9 kg vs. 44.4 kg; p = 0.0003). Absolute and relative liver weight was significantly (p < 0.0001) lower in flukicide-treated compared to untreated cattle. Overall, this analysis provided evidence of a substantial negative effect of early (migrating) liver fluke infection on the growth of young cattle, likely due to pathology of the liver and associated reduction in its function as the central organ for bioenergy and protein metabolism.

Plasma pharmacokinetics of clorsulon following administration of a single subcutaneous or intravenous injection to cattle.

The benzenedisulfonamide derivative clorsulon is a potent fasciolicide which is marketed in fixed combination injectables, typically combined with the macrocyclic lactone ivermectin. In the presented pharmacokinetic study, the plasma profile of clorsulon in 32 healthy, young Brown Swiss cattle was administered a single intravenous dose at 3 mg/kg body weight or subcutaneously at 3, 6 or 12 mg/kg body weight (4 intact male and 4 female animals per treatment) as a 30% w/v clorsulon injection formulation. Serial blood samples were collected up to 24 days after administration to establish the pharmacokinetics, bioavailability and dose proportionality of clorsulon. Following a single intravenous injection of clorsulon at 3 mg/kg body weight, the area under the concentration versus time curve from the start of dose administration to the time of the last quantifiable concentration (AUClast ) was 4830 ± 941 day*ng/mL, and half-live was 2.37 ± 0.98 days. The back extrapolated concentration at time 0 was 38,500 ± 6070 ng/mL. The volume of distribution at steady state and clearance were 685 ± 107 mL/kg and 664 ± 127 mL/day/kg, respectively. In the groups dosed at 3, 6 or 12 mg/kg body weight by subcutaneous injection, clorsulon plasma concentrations rose to maximum within 0.5 day and decreased to the last sample point. For these groups, the maximum plasma clorsulon concentrations were 3100 ± 838, 5250 ± 1220 and 10,800 ± 1730 ng/mL, respectively, and the AUClast was 5330 ± 925, 9630 ± 1300 and 21,500 ± 3320 day*ng/mL, respectively. Half-lives, 2.01 ± 0.62, 3.84 ± 1.42 and 5.36 ± 0.60 days, respectively, increased significantly with dose, likely related to increasing dose volume. Clorsulon was well absorbed and fully bioavailable (103%-114%) after subcutaneous injection. No gender-related difference in systemic exposure was observed. Assessment of Cmax and AUClast demonstrated a proportional increase in systemic exposure to the clorsulon subcutaneous doses over the range of 3-12 mg/kg body weight.

A note on the relationship between fecal larval excretion and Protostrongylus rufescens lungworm burden in sheep.

The examination of feces for stages of parasitic helminths is the most widely used methodology for the intravital diagnosis in domestic animals of patent endoparasitism including pulmonary nematode infections. Although there is only little information on the relationship of lungworm larval excretion and corresponding parasite burdens, fecal larval counts are used as indirect measure ("biomarker") for the intensity of infection, for instance in anthelmintic efficacy studies. To assess the relationship between fecal larval and Protostrongylus rufescens parasite counts in sheep, log-transformed data of 14 naturally infected animals were analyzed. The larval excretion of the sheep was monitored in approximately weekly intervals over 6 weeks before lungworm recovery. Analyses were performed on the larval counts (at a single time point or counts averaged over several consecutive time points) relating to parasite counts. Fecal larval counts and the P. rufescens nematode burden (range, 17 to 406) were significantly and strongly correlated (p < 0.05 for all analyses; Spearman's r > 0.6) with the number of larvae excreted increasing with increasing lungworm burden. Subsequently performed regression analysis demonstrated a statistically significant strong linear relationship between P. rufescens worm and fecal larval counts (p < 0.01 for all analyses; R2, range 0.5094 to 0.8150). Analyses based on larval counts averaged over two or more consecutive time points resulted in higher Spearman's r and R2 compared with analyses based on single time point larval counts. Despite of some variability, the analyses indicate that fecal larval counts can be regarded as a useful measure of the P. rufescens burden in sheep.

Pharmacokinetic-pharmacodynamic relationships of pour-on administered eprinomectin in nematode-infected lactating female and male castrated dairy breed goats.

Eprinomectin (EPM), a macrocyclic lactone with low excretion in milk and high efficacy against endoparasites and ectoparasites, is widely used in veterinary medicine. In this paper, EPM pharmacokinetics and anthelmintic efficacy previously established in one study with lactating female goats and three studies with male castrated growing dairy breed goats (all with induced mixed adult gastrointestinal nematode parasitism and treated with a single 1-mg/kg pour-on administration of EPM) were retrospectively evaluated using pharmacokinetic-pharmacodynamic (PK-PD) modeling. The PK-PD analyses between EPM exposure (Cmax and AUClast) and anthelmintic response (percent efficacy) were performed for lactating female goats only and pooled lactating female and male castrated goats. The Cmax and AUClast showed no significant difference between lactating female goats and combined male castrated goats. PK-PD modeling demonstrated Trichostrongylus colubriformis, a small-intestine nematode, as a suitable indicator of the EPM nematocidal efficacy. The EC90 values obtained by modeling Cmax vs T. colubriformis were 3.50 and 2.43 ng/mL for lactating female goats only and pooled lactating female and male castrated goats, respectively. The values of AUClast needed for 90% efficacy of T. colubriformis were 25.4 and 21.1 day*ng/mL for lactating female goats only and pooled lactating female and male castrated goats, respectively. Overall, the predicted pharmacological response against T. colubriformis is similar for lactating goats only and pooled lactating female and male castrated goats and correlates with observed efficacy. In conclusion, a dosage of 1-mg/kg EPM as a pour-on is sufficient to ensure efficacy against common nematodes in both lactating female and male castrated goats.

Naturally acquired ovine-adapted nematode infections in young cattle and their treatment with eprinomectin 5% w/v extended-release injection.

The objective of this controlled study was to determine the anthelmintic efficacy of eprinomectin 5% w/v extended-release injection (Eprinomectin ERI; LONGRANGE®, Boehringer Ingelheim Animal Health) against primarily ovine-adapted gastrointestinal strongylid nematode (GIN) parasites in naturally infected young cattle. Eighteen calves which grazed on a sheep-dominated mixed sheep-cattle farm were randomly allocated into two equal groups (saline [control] or Eprinomectin ERI, administered each at 1 mL/50 kg body weight once subcutaneously), treated and euthanized 14 days thereafter for a differential GIN count. Ten species of strongylid nematodes were recovered from the control calves (ovine-adapted Haemonchus contortus, Teladorsagia circumcincta, Trichostrongylus colubriformis, Cooperia curticei, Nematodirus battus, Chabertia ovina; bovine-adapted Ostertagia ostertagi, Cooperia oncophora, Nematodirus helvetianus; 'generalist' Trichostrongylus axei). Adult GIN counts ranged from 1,540 to 5,244 for the control calves and from zero to 110 for the Eprinomectin ERI-treated calves. Accepting the International Cooperation on Harmonisation of Technical Requirements for Registration of Veterinary Medicinal Products anthelmintic guidelines criteria on adequacy of infections to be demonstrated in the control animals, this study showed that Ch. ovina, C. curticei, H. contortus, N. battus, T. axei, and T. colubriformis were reduced significantly (p < 0.0001) by ≥ 98.7% in the animals treated with Eprinomectin ERI. In conclusion, Eprinomectin ERI treatment was efficacious against a range of ovine-adapted nematode parasites in naturally infected young cattle.

Distribution, prevalence, and intensity of Sarcocystis infections in sika deer (Cervus nippon) of free-ranging populations in Germany and Austria.

Sika deer (Cervus nippon), which are native to the Japanese islands and the adjacent mainland of eastern Asia, have been introduced into Europe and established free-ranging populations in several countries. Various Sarcocystis species have been identified recently from farmed "mainland sika" deer in Lithuania and native "Japanese sika" deer in Japan. To study the distribution, prevalence and intensity of Sarcocystis infection in free-ranging sika deer outside of their natural range heart and/or diaphragm samples of 311 animals from nine populations in Germany and Austria were examined by histology.Overall, sarcocysts were detected in either heart or diaphragm of 107/311 deer (34.4%) with prevalence ranging roughly from 5 to 50% among the populations. Considering the 263 animals with both heart and diaphragm available, prevalence varied significantly (p < 0.0001) among calves (20.2%), yearlings (40.3%), and adult deer (49.1%) but did not differ between male and female deer (48.3% vs. 43.7%; p = 0.6483). Occurrence of sarcocysts in heart vs. diaphragm indicated a marginal difference (27.8% vs. 20.9%; p = 0.0839). Intensity of infection in all but one heart positive and all diaphragm positive animals was low (< 10 sarcocysts per square centimeter muscle cut). While heart sarcocyst counts of yearlings and adult deer were higher than those of calves and were higher in male compared to female sika deer, diaphragm sarcocyst counts did not differ significantly between age groups and sexes. Sarcocystis infection was demonstrated at variable prevalence in sika deer in all populations but intensity is apparently low. Further studies are needed to identify the species of Sarcocystis infecting sika deer naturalized outside their natural range.

Eprinomectin 5% w/v extended-release injection for the treatment of larval and/or adult Bunostomum phlebotomum, Haemonchus contortus, H. placei, Nematodirus helvetianus, Oesophagostomum radiatum, and Trichostrongylus colubriformis infections in cattle.

In an effort to generate data for regulatory purposes on the therapeutic efficacy of eprinomectin 5% w/v extended-release injection (Eprinomectin ERI) (LongRange®, Boehringer Ingelheim) against infections of immature and adult stages of some nematode species (or stages) which are generally less common or predominantly seen in younger cattle, nine studies (minimum two per parasite and stage) were conducted in the USA and Germany. A total of 218 young cattle were included in seven experimentally induced infection studies (180 animals) and two studies with naturally acquired nematode infections (38 animals), which were compliant with WAAVP and VICH guidelines. In each study, cattle were randomly assigned into groups which received 1 mL per 50 kg body weight of either saline (controls) or Eprinomectin ERI (equivalent to 1 mg eprinomectin per kg body weight) via subcutaneous injection when the parasites were developing fourth-stage larval (L4) or adult nematodes. Following necropsy and parasite recovery, percentage efficacy was calculated based on the comparison of geometric mean nematode counts of the Eprinomectin ERI- vs. the saline-treated animals. Eprinomectin ERI-treated cattle had significantly (p < 0.05) lower counts of each species and stage of nematodes than the controls. Eprinomectin ERI treatment was demonstrated to be efficacious (> 90%) against L4 and adult Bunostomum phlebotomum and Nematodirus helvetianus; against L4 Haemonchus placei, Oesophagostomum radiatum, and Trichostrongylus colubriformis; and against adult H. contortus. Percentage efficacy against L4 H. contortus was variable (69.6 to 100%). All treatments were well accepted, and no treatment-related health problems were observed in any study.

Molecular identification of Sarcocystis species in diaphragm muscle tissue of European mouflon (Ovis gmelini musimon) from Austria.

Previous morphological studies suggested that mouflon may have sarcocysts similar to those of sheep. However, to date, no molecular-based studies of the species of Sarcocystis infecting mouflon have been done. The present study identified Sarcocystis species in diaphragm muscle samples from 20 European mouflon (Ovis gmelini musimon). Molecular identification using the cox1 sequence analysis was performed on sarcocysts excised from muscle tissue and on DNA from digested muscle samples. Both frequency and intensity of infection in mouflon were high with 19 of 20 animals testing Sarcocystis positive and > 50 cysts per gram of tissue recovered from 10 of the 19 Sarcocystis positive animals. Molecular analysis revealed dominant Sarcocystis tenella (18/19 animals) and Sarcocystis arieticanis (1/19 animals), whose known intermediate hosts are sheep. In addition, Sarcocystis capracanis, which is known to form sarcocysts in goats, was detected in two animals. The results of this study demonstrated the digestion method to be superior over the direct isolation of sarcocysts for the molecular identification of Sarcocystis species in a certain host. Future research of Sarcocystis diversity in wild ovine and caprine species is needed.

DNA barcoding of rumen flukes (Paramphistomidae) from bovines in Germany and Austria.

Rumen flukes have received growing veterinary attention in western and central Europe during the past two decades because of an increase in prevalence of infection in cattle and sheep, including cases of severe clinical disease. Historically, rumen fluke infections in Europe were assumed to be caused mainly by Paramphistomum cervi (or species, which were later considered to be synonymous with P. cervi), but more recently molecular studies demonstrated Calicophoron daubneyi to be the predominating species. For the present investigation, adult rumen flukes isolated from 23 cattle originating from ten farms in Germany (Saxony [1], Baden-Württemberg [4], Bavaria [5]) and one farm in Austria (Tyrol) were analyzed to establish partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI) and the complete sequence of the nuclear internal transcribed spacer 2 (ITS2). Flukes of five animals (dairy cows from three farms in Bavaria) were determined as P. leydeni, and flukes of 18 animals (dairy cows or cattle from cow-calf operations from eight farms in Saxony [1], Baden-Württemberg [4], Bavaria [2], and Tyrol [1]) were identified as C. daubneyi. Based on the molecular analysis of adult rumen flukes collected from cattle, the results of this investigation confirm the common occurrence of C. daubneyi in Germany and reveal the first definitive findings of P. leydeni in Germany and C. daubneyi in Austria.

Pour-on administration of eprinomectin to lactating dairy goats: Pharmacokinetics and anthelmintic efficacy.

Lactation is discussed as a physiological covariate which may influence the exposure characteristics of systemically acting drugs including macrocyclic lactones and potentially alter their pharmacological response. This study characterizes for the first time in the same study, the plasma profile and therapeutic anthelmintic efficacy of eprinomectin 5 mg/ml solution (EPRINEX® Multi, Boehringer Ingelheim) administered as a pour-on at 1 mg per kg body weight to lactating dairy goats. The study was conducted in compliance with VICH GCP and anthelmintic efficacy evaluation guidelines and included 20 goats harboring induced adult gastrointestinal and pulmonary nematode infections. The goats were blocked on pre-treatment body weight and randomly allocated either to remain untreated (control) or to be eprinomectin-treated. Plasma samples to determine the plasma disposition kinetics of eprinomectin (B1a component) were obtained at intervals up to 14 days following treatment when the animals were necropsied for parasite enumeration and identification. Basic pharmacokinetic parameters of eprinomectin determined in the ten eprinomectin-treated goats were as follows: AUClast , 23.8 ± 9.7 day*ng/ml and Cmax , 5.35 ± 2.27 ng/ml; individual maximum plasma concentrations were observed from 8 to 48 h after treatment (median Tmax , 0.5 days). Topical eprinomectin treatment efficacy, based on significant (p < .01) worm burden reductions in eprinomectin-treated animals relative to untreated controls, was ≥97% to 100% against adult Dictyocaulus filaria, Haemonchus contortus, Teladorsagia circumcincta(pinnata/trifurcata), Trichostrongylus axei, T. colubriformis, Cooperia curticei, Nematodirus battus, and Oesophagostomum venulosum. Both pharmacokinetic parameters and anthelmintic activity in lactating dairy goats were similar to those observed in parasitized young growing and adult female non-lactating dairy goats treated with eprinomectin administered as a pour-on.

Genetic diversity and phylogeography of the elusive, but epidemiologically important Echinococcus granulosus sensu stricto genotype G3.

Cystic echinococcosis (CE) is a severe parasitic disease caused by the species complex Echinococcus granulosus sensu lato. Human infections are most commonly associated with E. granulosus sensu stricto (s.s.), comprising genotypes G1 and G3. The objective of the current study was to provide first insight into the genetic diversity and phylogeography of genotype G3. Despite the epidemiological importance of the genotype, it has remained poorly explored due to the ambiguity in the definition of the genotype. However, it was recently demonstrated that long sequences of mitochondrial DNA (mtDNA) provide a reliable method to discriminate G1 and G3 from each other. Therefore, we sequenced near-complete mtDNA of 39 G3 samples, covering most of the known distribution range and host spectra of the genotype. The phylogenetic network revealed high genetic variation within E. granulosus s.s. G3 and while G3 is significantly less prevalent worldwide than G1, the genetic diversity of both of the genotypes is equally high. We also present the results of the Bayesian phylogeographic analysis, which yielded several well-supported diffusion routes of genotype G3 originating from Turkey and Iran, suggesting the Middle East as the origin of the genotype.

Anaplasma phagocytophilum and Babesia spp. in roe deer (Capreolus capreolus), fallow deer (Dama dama) and mouflon (Ovis musimon) in Germany.

Infections with the tick-borne pathogens Anaplasma phagocytophilum and Babesia spp. can cause febrile disease in several mammalian species, including humans. Wild ruminants in Europe are suggested to serve as reservoir hosts for particular strains or species of these pathogens. The aims of this study were to investigate the occurrence of A. phagocytophilum and Babesia spp. in roe deer (Capreolus capreolus), fallow deer (Dama dama) and mouflon (Ovis musimon orientalis) in Germany, and the diversity and host association of genetic variants of A. phagocytophilum and Babesia species. From 2009 to 2010, 364 spleen samples from 153 roe deer, 43 fallow deer and 168 mouflon from 13 locations in Germany were tested for DNA of A. phagocytophilum and Babesia spp. by real-time PCR or conventional PCR, respectively. Variants of A. phagocytophilum were investigated with a nested PCR targeting the partial 16S rRNA gene, and species of piroplasms were identified by sequencing. DNA of A. phagocytophilum was detected in 303 (83.2%) samples: roe deer, 96.1% (147/153); fallow deer, 72.1% (31/43); and mouflon, 74.4% (125/168). Sequence analysis of 16S rRNA-PCR products revealed the presence of nine different genetic variants. DNA of Babesia spp. was found in 113 (31.0%) samples: roe deer, 62.8% (96/153); fallow deer, 16.3% (6/43); and mouflon, 6.5% (11/168). Babesia capreoli, Babesia sp. EU1 (referred to also as B. venatorum), B. odocoilei-like and a Theileria species were identified. Co-infections with A. phagocytophilum and Babesia spp. were detected in 30.0% of the animals which were tested positive for A. phagocytophilum and/or Babesia spp. Roe deer had a significantly higher percentage of co-infections (60.8%), followed by fallow deer (14.0%) and mouflon (6.5%). Thus, the results suggest that roe deer plays a key role in the endemic cycles of the pathogens investigated.

Eprinomectin pour-on (EPRINEX® Pour-on, Merial): efficacy against gastrointestinal and pulmonary nematodes and pharmacokinetics in sheep.

BACKGROUND: The anthelmintic efficacy of the 0.5% w/v topical formulation of eprinomectin (EPN), EPRINEX® Pour-on (Merial) when administered at 1 mg/kg body weight was evaluated in sheep in two dose confirmation laboratory studies and one multicenter field study. In addition, the pharmacokinetics of EPN when administered at that dosage to adult sheep was determined. RESULTS: In the two dose confirmation studies, which included 10 sheep each, sheep treated with topical EPN had significantly (p < 0.05) fewer of the following nematodes than the untreated sheep with overall reduction of nematode counts by >99%: adult Dictyocaulus filaria, Haemonchus contortus, Teladorsagia circumcincta(pinnata/trifurcata), Trichostrongylus axei, T. colubriformis, T. vitrinus, Cooperia curticei, Nematodirus battus, Strongyloides papillosus, Chabertia ovina and Oesophagostomum venulosum, and inhibited fourth-stage Teladorsagia larvae. A total of 196 sheep harboring naturally acquired gastrointestinal nematode infections were included in the field efficacy study at two sites each in Germany (48 Merino x Ile de France lambs, 52 adult Merino females) and in Italy (adult male and female Bagnolese, Lacaune, Lacaune x Bagnolese, Bagnolese x Sarda sheep; 48 animals per site). Animals were blocked on pre-treatment body weight and within each block, one animal was randomly assigned to the control (untreated) group and three animals were randomly assigned to be treated with topical EPN. Examination of feces 14 days after treatment demonstrated that, relative to the controls, topical EPN-treated sheep had significantly (p < 0.0001) lower strongylid egg counts. Reduction was ≥97% at each site and 98.6% across all sites. Pharmacokinetics of EPN following single treatment with topical EPN were determined in eight ~4.5 year old female Merino cross sheep based on the analysis of plasma samples which were collected from two hours to 21 days following treatment. The main pharmacokinetic parameters were: Cmax 6.20 ± 1.71 ng/mL, AUClast 48.8 ± 19.2 day*ng/mL, Tmax 3.13 ± 2.99 days and T1/2 6.40 ± 2.95 days. No treatment-related health problems or adverse drug events were observed in any study. CONCLUSION: These studies demonstrated 0.5% w/v EPN administered topically at 1 mg/kg body weight to be highly efficacious against a broad range of ovine gastrointestinal nematodes and D. filaria lungworms and well tolerated by sheep of different ages, breeds, gender and physiological status.

Parasites and vector-borne diseases in client-owned dogs in Albania: infestation with arthropod ectoparasites.

To establish the diversity and seasonality of ectoparasite infestation in client-owned dogs in Albania, 602 dogs visiting four small animal clinics in Tirana from March 2010 to April 2011 inclusive were examined for ectoparasites by full body search and total body comb. In addition, ear swab specimens collected from all dogs and scrapings taken from skin lesions suspicious of mite infestation were examined for parasitic mites. Overall, 93 dogs (15.4 %, 95%CI 12.6-18.6) were demonstrated to be infested, and nine species of ectoparasites were identified: Ixodes ricinus, 0.8 %; Rhipicephalus sanguineus s. l., 8.1 %; Demodex canis, 0.2 %; Sarcoptes scabiei, 0.7 %; Otodectes cynotis, 2.8 %; Ctenocephalides canis, 4.8 %; Ctenocephalides felis, 3.0 %; Pulex irritans, 0.2 %; and Trichodectes canis, 0.2 %. Single and multiple infestations with up to four species of ectoparasites concurrently were recorded in 67 (11.1 %, 95%CI 8.7-13.9) and 26 dogs (4.3 %, 95%CI 2.8-6.3), respectively. On univariate analysis, the category of breed (pure breed dogs vs. mixed-breed dogs), the dog's purpose (pet, hunting dog, working dog), the housing environment (mainly indoors/indoors with regular outside walking vs. yard plus kennel/run), the history of ectoparasiticide treatment and the season of examination were identified as significant (p < 0.05) factors predisposing dogs to various ectoparasites, while the variables dog's age, gender, the dog's habitat (city, suburban, rural) and the presence/absence of other pets were not significant predictors. Multivariate logistic regression analysis for factors associated with overall ectoparasitism revealed that dogs treated with ectoparasiticides at least once per year (odds ratio [OR] = 0.24; p < 0.001) had a significantly lower risk of infestation compared with dogs not treated against ectoparasite infestation. Dogs examined during spring, summer and autumn (OR = 7.08, 7.43 and 2.48, respectively; all p < 0.001) had a significantly higher risk of infestation than dogs examined during winter. By providing basic data on the infestation with ectoparasites in client-owned, veterinary-cared-for dogs from Albania for the first time, the results of this survey should emphasize the need of an increase of attention to ectoparasites in dogs by both veterinarians and dog owners.

Efficacy against nematode infections and safety of afoxolaner plus milbemycin oxime chewable tablets in domestic dogs under field conditions in Europe.

Afoxolaner (AFX) plus milbemycin oxime (MO) combination chewable tablets (NexGard Spectra®, Merial) were evaluated for safety and efficacy against naturally acquired nematode infections in domestic dogs in a multi-centre, positive control, blinded field study using a randomized block design based on the order of presentation for allocation. In total, 408 dogs confirmed positive for naturally acquired infections of intestinal nematodes by pre-treatment faecal examination were studied in ten countries in Europe (Albania, Austria, Bulgaria, France, Germany, Hungary, Italy, Lithuania, Romania and Slovakia). Pre-treatment faecal examination revealed Toxocara, Toxascaris, hookworm, Trichuris and/or Capillaria nematode infections in 134, 30, 223, 155 and 14 dogs, respectively. Dogs were allocated to one of two treatment groups in a ratio of 1, AFX + MO chewables (≥2.5 mg AFX + ≥0.5 mg MO per kg body weight, according to dose bands; 207 dogs), and 1, MO plus praziquantel (PRZ) chewables (Milbemax®, Novartis; ≥0.5 mg MO + ≥5 mg PRZ per kg body weight, according to the manufacturer's instructions; 201 dogs) and treated once. For evaluation of efficacy based on reduction of faecal nematode egg counts, two faecal samples, one collected prior to treatment and one collected 9 to 21 days after treatment, were examined using modified McMaster techniques. For evaluation of systemic safety, dogs were examined by a veterinarian before treatment administration and at study end, and dog owners observed the health status of their dogs until the end of the study and reported any abnormal observation. For dogs treated with AFX + MO chewables, the efficacy was 99.7, 99.7, 97.2, 99.7 and 99.7 % for Toxocara, Toxascaris, hookworm, Trichuris and Capillaria, respectively; and the efficacy was 99.5, 99.4, 94.3, 99.9 and 98.0 %, respectively, for the MO + PRZ-treated dogs (p ≤ 0.002 for all nematodes and both treatments). For Toxocara, hookworm and Trichuris, non-inferiority analysis demonstrated that the efficacy of AFX + MO chewable tablets was equal to or better than that of MO + PRZ. In spite that both treatments were ≥98 % efficacious against Toxascaris and Capillaria, a hypothesis of non-inferiority for both genera could not be established due to the low number of dogs infected with these parasites. No treatment-related adverse experiences were observed throughout the study. For both treatments, all dogs were given a systemic safety score of 'excellent' apart from one dog in each treatment group which received a score of 'acceptable'. AFX + MO combination chewables were shown to be safe and demonstrated a high level of efficacy when administered once to dogs infected with a broad range of parasitic nematodes under field conditions.

High-resolution phylogeography of zoonotic tapeworm Echinococcus granulosus sensu stricto genotype G1 with an emphasis on its distribution in Turkey, Italy and Spain.

Echinococcus granulosus is the causative agent of cystic echinococcosis. The disease is a significant global public health concern and human infections are most commonly associated with E. granulosus sensu stricto (s. s.) genotype G1. The objectives of this study were to: (i) analyse the genetic variation and phylogeography of E. granulosus s. s. G1 in part of its main distribution range in Europe using 8274 bp of mtDNA; (ii) compare the results with those derived from previously used shorter mtDNA sequences and highlight the major differences. We sequenced a total of 91 E. granulosus s. s. G1 isolates from six different intermediate host species, including humans. The isolates originated from seven countries representing primarily Turkey, Italy and Spain. Few samples were also from Albania, Greece, Romania and from a patient originating from Algeria, but diagnosed in Finland. The analysed 91 sequences were divided into 83 haplotypes, revealing complex phylogeography and high genetic variation of E. granulosus s. s. G1 in Europe, particularly in the high-diversity domestication centre of western Asia. Comparisons with shorter mtDNA datasets revealed that 8274 bp sequences provided significantly higher phylogenetic resolution and thus more power to reveal the genetic relations between different haplotypes.

Control of parasitic infection with ivermectin long-acting injection (IVOMEC® GOLD) and production benefit in first-season grazing cattle facing a high-level larval challenge in Germany.

Gastrointestinal and pulmonary nematode infections are affecting the health and productivity of grazing cattle worldwide. To evaluate the effects of a single treatment with ivermectin long-acting injection (IVM LAI; IVOMEC® GOLD, Merial; 3.15 % ivermectin w/v) in first-grazing season cattle, two studies were conducted under continued stocking conditions for 84 or 100 days in Bavaria, Germany. Each study involved 68 naturally infected, approximately 4- to 6-month-old Brown Swiss bull calves. Animals were blocked based on pretreatment body weights. Within each block of four animals, animals were randomly assigned to treatments: one to saline (control) and three to IVM LAI. Treatments were injected at 1 mL/50 kg body weight subcutaneously in front of the shoulder. Animals in both studies were managed as one herd each grazing together. Cattle were weighed and fecal samples were collected pretreatment and at intervals thereafter for determination of weight gain and treatment efficacy, respectively. Fecal examination including composite fecal culture indicated the presence of nematodes of the genera Cooperia (dominating), Haemonchus, Nematodirus, Ostertagia, Strongyloides, Trichostrongylus, Trichuris, and Dictyocaulus, and Moniezia cestodes in the cattle. Following treatment, IVM LAI-treated cattle did not shed any Dictyocaulus larvae for 84 days while controls continued to pass larvae. Compared to the controls, IVM LAI-treated cattle had significantly (p < 0.01) lower strongylid egg counts at each occasion. Percentage reductions were ≥94 % up to 70 days after treatment and were ≥83.9 and 58.9 % at 84 and 100 days. Over the 84- or 100-day study periods, IVM LAI-treated cattle gained significantly more weight than the controls: 22.7 and 12.4 kg, respectively. The two studies demonstrated a high efficacy of IVM LAI against gastrointestinal and pulmonary nematode infections under field conditions in Germany which was associated with significant benefit as to weight gain.

Parasites and vector-borne pathogens in client-owned dogs in Albania. Blood pathogens and seroprevalences of parasitic and other infectious agents.

Knowledge on the epidemiology of parasitic and vector-borne infections is still very limited for Albania, a country located in the Balkan Peninsula in southeast Europe. Recent publications indicated prevalence rates of up to 52% for vector-borne infections in less-cared dogs in Albania. To provide data on the epidemiological situation in dogs under veterinary care, a total of 602 client-owned dogs presented to four small animal clinics between March 2010 and April 2011 in Tirana, Albania, were screened by examination of Giemsa-stained blood smears, PCR, and serological methods for the presence of arthropod-borne infections, as well as Neospora caninum and Toxoplasma gondii. Eight different pathogens, namely Babesia vogeli, Hepatozoon canis, Leishmania infantum, Dirofilaria immitis, Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, and Mycoplasma haemocanis, were detected by direct methods with prevalence rates ranging from 1 to 9%. Seroprevalence for Babesia spp., L. infantum, Anaplasma spp., and E. canis were 6.6, 5.1, 24.1, and 20.8%, respectively. Dogs >1 year of age were positive for vector-borne infections significantly more often than younger dogs (p = 0.003). More than half (51.7%) of the dogs were seroreactive to T. gondii and 18.3% to N. caninum. This is the first report on the detection of A. phagocytophilum, A. platys, E. canis, and M. haemocanis by PCR as well as the serological confirmation of exposure of dogs to N. caninum and T. gondii in Albania. The spectrum of pathogens and the seroprevalences for N. caninum and T. gondii in client-owned dogs from Tirana, Albania, are comparable to that reported in other countries in the Mediterranean Basin. The prevalence rates of vector-borne pathogens are at the lower range of that reported in studies from this geographical region. This is probably due to increased awareness of the owners of pet dogs, including better husbandry conditions and ectoparasiticidal treatment, thus limiting exposure of dogs to vectors.

Efficacy of oral afoxolaner plus milbemycin oxime chewables against induced infestations with Dermacentor reticulatus in dogs.

The efficacy of afoxolaner plus milbemycin oxime (AFX + MO) combination chewables (NexGard Spectra®, Merial) and AFX single-entity chewables (NexGard®, Merial) against induced infestations with Dermacentor reticulatus ticks was evaluated in dogs. Thirty dogs were assigned to blocks of three animals each based on pre-allocation tick counts and were randomly allocated to one of three groups: untreated (control), treated with a combination of AFX + MO chewables to be as close as possible to the minimum effective dose of AFX + MO (2.5 + 0.5 mg per kg body weight), and treated with a combination of NexGard® chewables to be as close as possible to the minimum effective dose of AFX (2.5 mg per kg body weight). Treatments were administered orally once on day 0. Starting 2 days before treatment administration, each dog was infested with approximately 50 ticks weekly for six consecutive weeks. Live ticks were counted at ∼48 h post-treatment (removal count) and at ∼48 h (in situ counts) and ∼72 h (removal counts) following each post-treatment infestation. Treatment with both AFX + MO and NexGard® chewables rapidly eliminated the existing tick infestations (100 % efficacy) within 2 days following treatment administration. Weekly re-infestations were controlled for a minimum of 5 weeks with the efficacy ranging from 92.2 to 99.7 % based on ∼48 h post-treatment in situ counts and between 99.0 and 100 % based on ∼72 h post-treatment removal counts (p < 0.0001 at each occasion). This study demonstrated a high efficacy of both AFX + MO chewable and NexGard® chewable treatments against infestations of dogs with D. reticulatus ticks for at least 5 weeks. In addition, this study indicated no interference between the two compounds with respect to the acaricidal activity provided by AFX.

Efficacy of Broadline against Capillaria aerophila lungworm infection in cats.

Capillaria aerophila is a globally distributed parasite of the respiratory system of carnivores and can be considered the second most common lungworm after Aelurostrongylus abstrusus in domestic cats in the northern hemisphere. To evaluate the efficacy of Broadline (Merial), a combination of fipronil 8.3% w/v, (S)-methoprene 10% w/v, eprinomectin 0.4% w/v and praziquantel 8.3% w/v against C. aerophila, a controlled study with 20 naturally infected cats was conducted. Following blocking by body weight, cats were allocated randomly to two groups of ten animals each: control (untreated) or treated once with Broadline according to the label instructions. For evaluation of efficacy, faeces were examined for capillarid egg shedding prior to and weekly for 3 weeks after treatment, when the cats were necropsied for C. aerophila recovery. Following single topical administration of Broadline, faecal capillarid egg counts were significantly reduced by 93.5 to 99.1% (p < 0.01) compared to the controls. Cats treated harboured significantly fewer C. aerophila lungworms compared to the untreated controls (efficacy 82.4%, p = 0.016). Results of this study demonstrate that Capillaria lungworm burdens can be markedly reduced and that faecal egg shedding can be substantially lowered or eliminated following a single treatment with Broadline.