Steffen Rehbein

Genetic Variants of Anaplasma phagocytophilum in Wild Caprine and Cervid Ungulates from the Alps in Tyrol, Austria

The occurrence of genetic variants of Anaplasma phagocytophilum was studied in wild ungulates from the northern and central eastern Alps in Tyrol, Austria. For this purpose, spleen samples collected from 53 game animals during the hunting season 2008/2009 (16 roe deer [Capreolus capreolus], 10 red deer [Cervus elaphus], 16 Alpine chamois [Rupicapra r. rupicapra], 7 Alpine ibex [Capra i. ibex], and 4 European mouflons [Ovis orientalis musimon]) were analyzed. Thirty-five animals originated from the Karwendel mountains, 12 from the Kaunertal area (Ötztal Alps), and the remaining from other mountainous areas in Tyrol. DNA extracts were screened with a real-time polymerase chain reaction targeting the msp2 gene of A. phagocytophilum. A total of 23 (43.4%) samples, from all ungulate species studied, were A. phagocytophilum positive. As of the date of this article, A. phagocytophilum has not been reported in the Alpine ibex. The positive samples were investigated further with polymerase chain reactions for amplification of the partial 16S rRNA, groEL, and msp4 genes. Sequence analysis using forward and reverse primers revealed seven different 16S rRNA gene variants. No variant could be attributed to any particular ungulate species. The groEL gene revealed 11 different variants, which grouped in the phylogenetic analysis into two distinct clusters: one cluster contained the sequences from roe deer, whereas the sequences of the other species formed the second cluster. The msp4 gene showed a high degree of variability in the amplified part with a total of 10 different sequence types. The results show that the wild mountain ungulates were infected to a considerable extent with various variants of A. phagocytophilum. The pathogenicity of the variants and the reservoir competence of the species investigated in this study deserve further attention in future studies.

Parasitic infections of domestic cats, Felis catus, in western Hungary.

During 2011, faeces from 235 owned domestic cats from a rural area in western Hungary were examined using standard coproscopical techniques. The overall prevalence of cats with endoparasites was 39.6% (95% CI 33.3-46.1). The most frequently identified faecal forms were those of ascarids (Toxocara, 17.4%; Toxascaris 7.2%), followed by those of Aelurostrongylus lungworms (14.5%), hookworms (11.1%), taeniid cestodes (4.7%), Cystoisospora coccidians (4.3%), and capillarids (3.8%). Single and multiple infections with up to five parasites concurrently were founded in 24.7% and 14.9% of the cats, respectively. Mixed endoparasite infections were recorded more frequently (p=0.0245) in cats greater than one year old compared to younger cats. Young cats (≤ 1 year) were parasitized more frequently (p<0.05) with ascarids and Cystoisospora spp. but demonstrated infections of hookworms, lungworms and taeniid cestodes less often than the older cats. Cats with taeniid infection were more likely (p<0.05) to harbour Toxocara, hookworm, Aelurostrongylus, and capillarid infections than cats without taeniid cestodes. Cats of owners who claimed the use of wormers were less frequently helminth-positive compared to cats whose owners did not use anthelmintics (21.2% vs. 44.4%; p=0.001). A subset of 115 faecal samples screened by a coproantigen ELISA revealed Giardia-specific antigen in 37.4% samples. Giardia cysts were found by immunofluorescent staining in 30 of the 43 samples tested positive for Giardia by ELISA. In addition, ectoparasites collected from 82 cats by body search and combing were identified. Fleas (1-30 per cat), biting lice (Felicola subrostratus), and ticks (1-5 per cat) were isolated from 58, 1 and 43 cats, respectively. Ctenocephalides felis was identified on all flea infested cats while single specimens of C. canis and Pulex irritans were recovered from three and two cats, respectively. All but one tick collected were adult Ixodes ricinus; the single other tick was a nymph of I. canisuga. By providing basic data on the epidemiology of parasitic infections, the results of this survey should emphasize the need of attending to parasites of cats from the veterinary point of view with respect to both appropriate diagnostics and control.

Re-evaluation of ivermectin efficacy against equine gastrointestinal parasites

Two trials were conducted to confirm the efficacy of ivermectin paste against endoparasites of horses. In these trials, 20 ponies were treated with ivermectin oral paste at 200 mcg x kg body weight once on Day 0, and 20 ponies served as unmedicated controls. The animals carried naturally acquired parasite infections as confirmed by pretrial fecal examination. The animals were necropsied for worm recovery on Days 14, 15 or 16. Parasites recovered were identified to species. Horses treated with ivermectin had significantly (P<0.05) fewer (>99.0% reduction) adult small strongyles (Coronocyclus spp including C. coronatus, C. labiatus, C. labratus; Cyathostomum spp including C. catinatum, C. pateratum; Cylicocyclus spp including C. ashworthi, C. elongatus, C. insigne, C. leptostomum, C. nassatus, C. radiatus; Cylicodontophorus bicoronatus; Cylicostephanus spp including C. asymetricus, C. bidentatus, C. calicatus, C. goldi, C. longibursatus, C. minutus; Gyalocephalus capitatus; Parapoteriostomum spp including P. euproctus, P. mettami; Petrovinema poculatum; Poteriostomum spp including P. imparidentatum, P. ratzii) and adult large strongyles (Strongylus edentatus, S. vulgaris; Triodontophorus spp including T. brevicauda, T. serratus; Craterostomum acuticaudatum) than the controls. Ivermectin was also highly effective (94% to >99%, P<0.05-0.01) against Gasterophilus intestinalis larvae, Habronema spp., Oxyuris equi, Parascaris equorum. The data from these two trials confirm that ivermectin paste administered to horses orally at 200mcg x kg(-1) continues to be highly effective for treatment and control of a broad range of small and large strongyle species as well as other species of gastrointestinal parasites.

Endoparasites of cats from the Tirana area and the first report on Aelurostrongylus abstrusus (Railliet, 1898) in Albania

ZusammenfassungIm Kot von Katzen aus dem Großraum Tirana wurden Larven nachgewiesen, die für eine Infektion mit dem Lungenwurm Aelurostrongylus abstrusus sprachen. Dieser Befund konnte durch die Sektion von 18 Katzen bestätigt werden, von denen 9 einen Befall der Lungen mit adulten A. abstrusus (Railliet, 1898) (Prävalenz 50 %; 1–11 Würmer) aufwiesen. Neben A. abstrusus wurden Eucoleus aerophilus (16,7 %; 1–9 Würmer) aus der Lunge sowie Toxocara cati (83,3 %; 2–33 Würmer), Ancylostoma tubaeforme (44,4 %; 1–20 Würmer), Dipylidium caninum (83,3 %; 1–164 Würmer), Joyeuxiella pasqualei (11,1 %; 1–3 Würmer) und ein Kratzer (Acanthozephala) aus dem Magen–Darm–Trakt der Katzen isoliert. Im Rektumkot waren Oozysten von Cystoisospora felis und C. rivolta bei 5,6 % bzw. 11,1 % der sezierten Katzen nachweisbar. Die vorläufigen Ergebnis sprechen für eine hohe Prävalenz an Endoparasiten in den freilebenden Katzen aus Tirana. Das Vorkommen von A. abstrusus, dessen Befall mit respiratorischen Symptomen einhergehen kann, wird hiermit erstmalig in Albanien beschrieben.SummaryFollowing the recovery of first–stage nematode larvae indicative of Aelurostrongylus abstrusus infection in the faeces of free-roaming cats from the greater Tirana area, examination of 18 cats at necropsy revealed nine of them harbouring adult A. abstrusus (Railliet, 1898) in the lungs (prevalence, 50%; range, 1–11). In addition to A. abstrusus, Eucoleus aerophilus (16.7%; 1–9) was isolated from the lungs, and Toxocara cati (83.3%; 2–33), Ancylostoma tubaeforme (44.4%; 1–20), Dipylidium caninum (83.3%; 1–164), Joyeuxiella pasqualei (11.1%; 1–3) and one specimen of an acanthocephalan (5.5%) were recovered from the gastrointestinal tract. Furthermore, oocysts of Cystoisospora felis and C. rivolta were found in the rectal faeces of 5.6% and 11.1% of the cats, respectively. In conclusion, the prevalence of endoparasite infection in free–roaming cats in Tirana can be considered to be high. The occurrence of A. abstrusus, which may cause respiratory distress in cats, is reported for the first time in Albania.

Detection of Babesia canis subspecies and other arthropod-borne diseases in dogs from Tirana, Albania

ZusammenfassungDurch Arthropoden übertragene Infektionen haben in der jüngeren Vergangenheit zunehmend an Bedeutung gewonnen, auch bedingt durch vermehrte Reisen in, beziehungsweise Importe von Hunden aus Regionen, in denen die Erreger endemisch sind. Während die epidemiologische Situation im westlichen Mittelmeerraum gut dokumentiert ist, sind aus Osteuropa und dem Balkan vergleichsweise wenige Informationen verfügbar. In der vorliegenden Studie wurden Blutproben von 30 klinisch unauffälligen Hunden aus den Randgebieten von Tirana, Albanien, auf vektor-übertragene Infektionen untersucht. Mittels direkter und/oder indirekter Verfahren wurden die Blutproben auf Babesia canis, Hepatozoon spp., Leishmania spp., Dirofilaria spp., Ehrlichia canis, Anaplasma phagocytophilum, Bartonella spp. und Rickettsia spp. untersucht. Im Blut von 20 Hunden (= 67 %) wurden Antikörper bzw. Erreger durch Arthropoden übertragene Infektionen nachgewiesen. Antikörper gegen B. canis, E. canis und/oder A. phagocytophilum waren im Serum von 19 Hunden (= 63 %) nachweisbar. Bei 13 Hunden (= 43 %) erfolgte ein Erregernachweis mittels Blutausstrich, PCR oder ELISA, wobei B. caniscanis, B. canis vogeli, Hepatozoon spp., D. immitis und/oder E. canis identifiziert wurden. Infektionen mit Leishmania spp., Bartonella spp. und Rickettsia spp. waren nicht nachweisbar.SummarySummary. The importance of arthropod-borne diseases increased in the recent past in particular due to frequent travel with dogs in or by importing of dogs from regions with endemic occurrence of these diseases. While the epidemiological situation is well known for the western parts of the Mediterranean, only limited data is available for Eastern Europe and the Balkans. Thirty clinically healthy dogs from suburban areas of Tirana, Albania, were tested for Babesia canis, Hepatozoon spp., Leishmania spp., Dirofilaria spp., Ehrlichia canis, Anaplasma phagocytophilum, Bartonella spp. and Rickettsia spp. using direct and indirect methods. Antibodies against and/or pathogens of arthropod-borne diseases were detected in the blood of 20 (67%) dogs. Nineteen dogs (63%) had antibodies against B. canis, E. canis and/or A. phagocytophilum.Babesia c. canis, Babesia c. vogeli, Hepatozoon spp., D. immitis and/or E. canis were identified by blood smear, PCR or ELISA in 13 (43%) dogs. There was no evidence for Leishmania spp., Bartonella spp. and Rickettsia spp. infections.

Persistent efficacy of topical eprinomectin against nematode parasites in cattle

Abstract Six studies were conducted to evaluate the persistent efficacy of eprinomectin pour-on against experimental challenges with infective nematode larvae in calves. In each study, calves were randomly assigned to one untreated group and up to four test groups, which were treated with eprinomectin at 500 μg/kg body weight at weekly intervals before single bolus challenge. The calves were necropsied approximately 4 weeks after challenge infection for nematode recovery. Eprinomectin pour-on provided ≥90% efficacy against challenge with Haemonchus placei, Trichostrongylus axei and T. colubriformis at 21 days after treatment and against Cooperia oncophora, C. punctata, C. surnabada, Dictyocaulus viviparus, Nematodirus helvetianus, Oesophagostomum radiatum and Ostertagia ostertagi at 28 days after treatment.

The efficacy of eprinomectin against induced infections of immature (fourth larval stage) and adult nematode parasites in cattle

Eprinomectin is a new endectocide of the avermectin chemical group developed for use in cattle. To establish its effectiveness against Cooperia spp., Dictyocaulus viviparus, Haemonchus contortus, Nematodirus helvetianus, Oesophagostomum radiatum, Ostertagia ostertagi, Trichostrongylus axei and Trichostrongylus colubriformis (immature infections only), six trials were conducted in Europe at two trial sites. In each trial, animals were artificially infected with a number of nematode species and treated with eprinomectin. In three trials, the nematodes were expected to be at the fourth larval stage of development at the time of treatment, and at the adult stage at the time of treatment in the other three trials. When compared with the untreated control groups, efficacy against the adult and immature nematode infections was > 99% for all the nematode species examined. There was no difference in efficacy between groups of animals with clipped hair at the application site, and those with long hair. Subsequent examination of the application sites, both visual and by palpation, showed no gross reactions to the applied treatments.

Survey of Toxoplasma gondii and Neospora caninum , haemotropic mycoplasmas and other arthropod-borne pathogens in cats from Albania

BackgroundAlbania is a country on the western part of the Balkan Peninsula. The Mediterranean climate is favourable for the stable development of many arthropod species, which are incriminated as vectors for various agents. Recently, several papers have reported on epidemiological aspects of parasitic diseases including vector-borne disease agents of dogs with zoonotic characteristics in Albania. However, data on the epidemiology of feline parasitic and bacterial agents in Albania is scarce.MethodsSerum and EDTA-blood samples collected from 146 domestic cats from Tirana during 2008 through 2010 were examined for exposure to Toxoplasma gondii, Neospora caninum, Leishmania infantum, and Anaplasma spp. with IFAT, for infection with L. infantum, A. phagocytophilum, Bartonella spp. and haemotropic mycoplasmas with conventional PCR and real-time PCR and for Dirofilaria immitis with antigen ELISA. Additionally blood smear microscopy was carried out for detection of blood-borne pathogens.ResultsAntibodies to T. gondii (titre ≥1:100) were demonstrated in 91 cats (62.3%). Antibodies to N. caninum (titre ≥1:100), L. infantum (titre ≥1:64) and Anaplasma spp. (titre ≥1:100) were found in the serum of 15 (10.3%), 1 (0.7%) or 3 (2.1%) cats, respectively. DNA of haemotropic mycoplasmas was detected in the blood of 45 cats (30.8%), namely Candidatus Mycoplasma haemominutum (21.9%), Mycoplasma haemofelis (10.3%), and Candidatus Mycoplasma turicensis (5.5%), with ten cats harbouring co-infections of two mycoplasmas each; blood from one cat was PCR positive for Bartonella henselae. No DNA of Leishmania spp. and A. phagocytophilum or circulating D. immitis antigen was detected in any cat sample. The overall prevalence of haemotropic mycoplasmas was significantly higher in male compared to female cats (40.6% vs. 24.1%, p = 0.0444); and age was associated positively with the prevalence of antibodies to T. gondii (p = 0.0008) and the percentage of haemotropic mycoplasma infection (p = 0.0454).ConclusionsWith the broad screening panel including direct and indirect methods applied in the present study, a wide spectrum of exposure to or infection with parasitic or bacterial agents was detected.

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.

Occurrence of Dipylidium caninum in fleas from client-owned cats and dogs in Europe using a new PCR detection assay

Ctenocephalides fleas are not only the most prevalent ectoparasites of dogs and cats but also the intermediate host of the cestode Dipylidium caninum. Due to the poor sensitivity of coproscopy to diagnose cat and dog infestation by Dipylidium, few epidemiological data are available on its prevalence among pet populations. A new PCR method was developed to specifically identify D. caninum rDNA inside single fleas. The PCR test was then applied to 5529 fleas of Ctenocephalides genus, 2701 Ctenocephalides felis fleas (1969 collected on 435 cats and 732 on 178 dogs) and 2828 Ctenocephalides canis fleas collected from 396 dogs. Precisely, 4.37% of cats were infested by a flea population infected with D. caninum. Out of the 1969 C. felis from cats, 2.23% were found to be infected with Dipylidium. From the 396 dogs infested with C. canis, 9.1%% were infested with the Dipylidium infected fleas, which is significantly higher than the observation made in cats (p=0.03). Moreover, 3.1% of the C. canis fleas were found to be infected with Dipylidium, which is not significantly different than in C. felis. Looking at the number of infected fleas in the positive samples (at least one PCR positive flea in a sample), the infestation rate in samples was varied from 3 to 100% with an average of 19.7% which is in favour of easy and regular Dipylidium reinfestations of both cats and dogs in households. For the first time, the spread of D. caninum between fleas and dogs and cats is confirmed throughout Europe.