Co-infection of Trypanosoma pestanai and Anaplasma phagocytophilum in a dog from Germany.

Infections with arthropod-borne pathogens are an increasing threat world-wide that requires heightened vigilance from veterinary and medical practitioners, especially when they involve new or unusual organisms. A dog was presented to a local veterinary clinic in Germany with malaise, pale mucous membranes and stiff joints. Clinical assessment revealed pyrexia, leukopenia and thrombocytopenia. On suspicion of a tick-borne infection, blood samples were examined for clinical and biochemical parameters and subjected to a Anaplasma phagocytophilum-, Borrelia spp.- and Ehrlichia canis-specific real-time PCR. Additionally, a sample of the pre-therapeutic buffy coat was co-cultured with the Ixodes scapularis cell-line ISE6 for 20days. Only the PCR specific for A. phagocytophilum DNA yielded a positive result, and furthermore, Anaplasma morulae were visible in granulocytes and tick cells. After co-culturing, extracellular trypomastigote and epimastigote stages of Trypanosoma sp. with an average length of 29.7µm were observed, featuring a pointed posterior end. Sequence analysis of a 2080bp fragment of the 18S rRNA gene showed 99% identity to the 18S rRNA gene of Trypanosoma pestanai, previously described from a European badger (Meles meles) in France. The dog's condition improved rapidly in response to doxycycline treatment for three weeks. The clinical status normalized and clinical blood parameters were found to be within the reference ranges. To our knowledge this is the first description of T. pestanai infection in a dog, the first detection of T. pestanai in Germany and the first documented co-infection with these two pathogens. Co-infections with unusual opportunistic vector-borne pathogens should be considered, if acute canine granulocytic anaplasmosis is evident.

[Nematode infections of the respiratory tract in dogs in Germany]. / Nematodeninfektionen des Respirationstrakts bei Hunden in Deutschland.

Recent studies have shown that the risk of lungworm infection may have increased in dogs in Germany in recent years. Analysis of the fecal examination of dogs has shown that Angiostrongylus vasorum, Crenosoma vulpis and Eucoleus aerophilus are endemic in Germany. Infections with A. vasorum were diagnosed in 223 of the examined dogs. A total of 102 A. vasorum-positive dogs were located in Baden-Wuerttemberg, 65 in North Rhine-Westphalia, 27 in Saarland, 15 in Bavaria, 7 in Rhineland-Palatinate, 5 in Hessen and 2 in Brandenburg. A total of 170 dogs were infected with C. vulpis, 54 of which came from North Rhine-Westphalia, 40 from Baden-Wuerttemberg, 30 from Bavaria, 17 from Rhineland-Palatinate, 17 from Saarland, 9 from Hessen and 1 each from Lower Saxony, Berlin and Mecklenburg-Western Pomerania. Infections with E. aerophilus were detected in 83 dogs, of which 23 lived in Baden-Wuerttemberg, 20 in North Rhine-Westphalia, 17 in Bavaria, 11 in Rhineland-Palatinate, 7 in Hessen, 4 in Saarland and 1 in Mecklenburg-Western Pomerania. Lungworm infections in dogs appear to be well established in Germany. The aim of the study presented was to assess the main facts, occurrence, geographical distribution, clinical signs, diagnosis and therapy of lungworm infections in dogs.

Detection of specific antibodies in dogs infected with Angiostrongylus vasorum.

Canine angiostrongylosis, caused by the nematode Angiostrongylus vasorum, is an emerging cardiopulmonary disease in Europe which can be fatal if left untreated. We determined the diagnostic value of the specific detection of antibodies against A. vasorum adult somatic antigen, adult excretory/secretory (E/S) antigen and first stage larvae (L1) somatic antigen in ELISAs. Also, A. vasorum adult somatic antigen purified by monoclonal antibodies (mAb) was evaluated in a sandwich-ELISA. Among the crude antigens, the best sensitivities when testing 21 naturally infected dogs were obtained using adult E/S and somatic antigen (85.7% and 76.2%, respectively), which were comparable with the results of the sandwich-ELISA based on mAb-purified antigens (81%). The ELISA performed with L1 antigen had the lowest sensitivity (42.9%). In experimentally inoculated dogs, the sensitivities ranged from 97.7% to 100% with all test settings. The specificity was 98.8% (92.5-99.9%, 95% CI) with all ELISAs using sera of 82 randomly selected dogs. Cross-reactions using adult somatic, adult E/S and L1 somatic antigen were observed in sera of dogs infected with Crenosoma vulpis, Dirofilaria immitis, Dirofilaria repens, and Eucoleus aerophilus. In contrast, using the mAb-purified antigens, the cross-reactions were minimal. Depending on the antigens used, specific antibodies were detected starting between 13 and 21 days post experimental inoculation (dpi), and at latest between 35 and 48 dpi, thus before or around the onset of patency. The serological follow-up of four A. vasorum-infected dogs after anthelmintic treatment at 88 dpi showed a decrease of antibody levels after drug administration, and the animals became seronegative 2-9 weeks later. Two untreated dogs remained seropositive. In four dogs treated 4 dpi, virtually no antibody-reaction was detectable, with the exception of the ELISA performed with L1 antigen. The early detection of specific antibodies against A. vasorum by ELISA represents a valid alternative for a reliable diagnosis and for follow-up investigations after anthelmintic treatment.

An ELISA for sensitive and specific detection of circulating antigen of Angiostrongylus vasorum in serum samples of naturally and experimentally infected dogs.

Canine angiostrongylosis is an emerging cardiopulmonary disease in Europe which can be fatal if left untreated. We developed a sandwich-ELISA based on a monoclonal antibody (mAb Av 56/1/2) and on polyclonal rabbit antibodies directed against Angiostrongylus vasorum adult excretory/secretory - antigen for the detection of circulating serum antigen of A. vasorum. The sensitivity of the test was 95.7% (78.1-99.9, 95% CI) as determined with sera of 23 dogs naturally infected with A. vasorum. The specificity was 94.0% (83.5-98.7, 95% CI) using 50 dog sera (control group) submitted for reasons other than parasitic infections. Potential cross-reactions were investigated with sera of a group of totally 61 dogs with proven infections with Dirofilaria immitis (n=23), Crenosoma vulpis (n=14), Ancylostoma caninum (n=4) or Toxocara canis (n=20). No significant difference was observed concerning the proportion of positive reactions between the control group and the group with proven helminth infections other than A. vasorum. In experimentally inoculated dogs with proven worm burdens of A. vasorum, the proportion of seropositive dogs increased over the first 3 months of infection, starting from 35 days post inoculation (dpi) which was before the onset of larval excretion. Ten weeks post inoculation, 98.6% of the dogs were seropositive, and circulating antigen persisted in two dogs with long-term follow-up over 286 and 356 days, respectively. In contrast, in dogs with a single treatment with imidacloprid/moxidectin at four or 32 dpi, no circulating antigen was observed, while in dogs treated at 88-92 dpi, OD values decreased within 13-34 days. The specific detection of circulating A. vasorum antigen by ELISA represents a valid alternative for reliable diagnosis and for follow-up investigations after anthelmintic treatment. Moreover, the test can be used for mass screening in large epidemiological investigations.

Atypical Toxoplasma gondii genotypes identified in oocysts shed by cats in Germany.

A total of 18,259 feline faecal samples from cats in Germany were collected and analysed for the presence of Toxoplasma gondii oocysts between June 2007 and December 2008. The proportion of T. gondii-positive samples collected between January and June was significantly lower than between July and December. The age of cats shedding T. gondii oocysts was not significantly different from the age of negative control cats. Forty-six T. gondii-positive samples were genetically characterised using nine PCR-restriction fragment length polymorphism (RFLP) markers which included newSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. In addition, 22 isolates that had already been partially characterised in a previous study were further typed using PCR-RFLP markers c22-8, c29-2, L358, PK1 and Apico. Genotyping of the 68 isolates revealed that the majority of T. gondii isolates (n=54) had Type II patterns at all loci but displayed a Type I pattern at the Apico locus. Three isolates displayed Type II patterns at all loci, including the Apico locus. In addition, we detected one isolate with clonal Type III patterns at all loci and three isolates with atypical and mixed genotypes. Seven isolates could not be fully genotyped. One of those isolates displayed alleles of both Types I and II at the Apico locus. To our knowledge this is the first description of the presence of T. gondii genotypes different from the clonal Types I, II and III in the faeces of naturally infected cats.

Oocysts of Neospora caninum, Hammondia heydorni, Toxoplasma gondii and Hammondia hammondi in faeces collected from dogs in Germany.

Faecal samples of 24,089 dogs were examined coproscopically in two veterinary laboratories in Germany between March 2001 and October 2004. In 47 dogs, oocysts of 9-14 microm size were found. Their morphology was similar to those of Hammondia heydorni and Neospora caninum. Samples of 28 of these dogs were further examined by inoculation into gerbils: seven isolates induced a specific antibody response against antigens of N. caninum NC-1 tachyzoites. This response suggests that the isolates contained N. caninum. In addition to H. heydorni (12 times isolated), Toxoplasma gondii occysts (twice) and Hammondia hammondi oocysts (twice) were observed in dog faeces. The latter findings suggest that coprophagia with a subsequent intestinal passage by dogs plays a role in the dissemination of coccidian parasites for which cats are definitive hosts. Five of the seven N. caninum (NC-GER2, NC-GER3, NC-GER4, NC-GER5, NC-GER6) and the two T. gondii isolates (TG-dgGER1, TG-dgGER2) were successfully passaged into cell culture and are now available for detailed characterization. In contrast to oocysts of other parasites, N. caninum oocysts were predominantly found between January and April (Fisher exact; P=0.038). In the sera of dogs shedding N. caninum, no reactions against the immunodominant antigens with apparent molecular weights of 19, 29, 30, 33 and 37 kDa of N. caninum tachyzoites were observed 3-5 weeks after shedding. However, the animals recognized a 152-kDa N. caninum antigen. Compared with those identified as H. heydorni, T. gondii or H. hammondi, N. caninum oocyst isolates were significantly smaller in length with the 75th percentiles

Endoparasites in dogs and cats in Germany 1999-2002.

Infections with endoparasites in dogs and cats have been determined by analysing the results of faecal examinations (Flotation, MIFC, sedimentation, Baermann, smear, ProSpecT Giardia Microplate Assay). Samples of 8438 dogs and 3167 cats from the years 1999 until 2002 have been included in the investigation. 2717 dogs (32.2%) and 771 cats (24.3%) have been infected with endoparasites. In the infected dogs the following parasites have been identified: Class Nematodea: Toxocara canis: 22.4%, Toxascaris leonina: 1.8%, Ancylostomatidae: 8.6%, Trichuris vulpis: 4.0%, Capillaria spp.: 2.3%, Crenosoma vulpis: 0.9%, Angiostrongylus vasorum: 0.3%; Class Cestodea: Taeniidae: 1.2%, Dipylidium caninum: 0.4%, Diplopylidium/Joyeuxiella: 0.1%, Mesocestoides: 0.2%, Diphyllobothrium latum: < 0.1%; Class Sporozoea: Sarcocystis spp.: 9.0%, Cystoisospora spp.: 22.3%, C. canis: 8.0%, C. ohioensis: 17.0%, Hammondia/Neospora: 1.7%; Class Zoomastigophorea: Giardia spp.: 51.6%. In the 771 infected cats the following prevalences of parasites have been found: Class Nematodea: Toxocara mystax: 26.2%, Ancylostoma tubaeforme: 0.3%, Capillaria spp.: 7.0%, Aelurostrongylus abstrusus: 2.7%; Class Cestodea: Taeniidae: 2.6%, Dipylidium caninum: 0.1%; Class Sporozoea: Sarcocystis spp.: 2.2%, Cystoisospora spp.: 21.9%, C. felis: 15.3%, C. rivolta: 7.9%, Toxoplasma/Hammondia: 4.5%; Class Zoomastigophorea: Giardia spp.: 51.6%.

[Migration and transposition of Oesophagostomum quadrispinulatum larvae from feces]. / Untersuchungen zur Migration und Translation von Oesophagostomum-quadrispinulatum-Larven aus dem Kot.

Investigations on the migration and translation of free-living stages of Oesophagostomum quadrispinulatum, using faeces containing eggs as starting material, revealed that mostly third stage larvae and very few second stage larvae migrated out of faeces, whereas first stage larvae remained in the faeces. The emigration rates depended on ambient relative humidity. Compared with the control, third stage larvae emigrated out of faeces at rates of 0.3%, 1.6% and 12.2% at 50%, 75% and 100% relative humidity, respectively. Offering helminth-free faeces, emigrated third stage larvae returned into faces at rates of 0.4-1.2%, 5.8-17.7%, 39.0-52.7%, and 45.2-60.7% after 1 h, 24 h, 5 days and 14 days, respectively. After a period of 1, 2, 3 or 4 weeks staying out of faeces, emigrated third stage larvae returned into faeces at rates of 23.3%, 8.8%, 22.1% and 6.0%, respectively. An examination of the horizontal translation revealed that most of the third stage larvae migrated distances up to 80 cm and a few even up to 150 cm returning into helminth-free faeces.

[The effect of optical radiation on the migration of third stage larvae of Oesophagostomum quadrispinulatum out of feces]. / Zum Einfluss optischer Strahlung auf die Emigration der Drittlarven von Oesophagostomum quadrispinulatum aus dem Kot.

The influence of optical radiation on the emigration of third stage larvae out of faeces was investigated by exposing faeces containing infective larvae of Oesophagostomum quadrispinulatum to broad-band radiation of a sun-simulating wavelength spectrum corresponding to a sunny day, a cloudy day, dawn, and a full-moon night, as well as to monochromatic radiation of different wavelength spectrum at dawn and to complete darkness. It was demonstrated that third stage larvae of O. quadrispinulatum were able to differentiate between daylight and darkness responding to different irradiances with very high emigration rates at irradiances corresponding to dawn, and significantly lower emigration rates corresponding to full-moon light, and darkness as well as a cloudy and sunny day. Infective larvae reacted to monochromatic radiation of different wave-length spectrum at dawn and showed significantly higher emigration rates at the violet, green, yellow, and red light wavelength compared to darkness.

Ectopic enterobiasis: a case report and review.

Enterobius vermicularis ('pinworm') is rarely found outside the gastro-intestinal tract. We describe a case of extra-intestinal pinworm abscess associated with an inguinal hernia in an adult. A brief review of the literature is given and possible mechanisms of tissue invasion are discussed.

[Helminth infections in wild boars kept in enclosures in southern Germany: severity of infections and fecal intensity]. / Helmintheninfektionen von Wildschweinen bei Gatterhaltung in Süddeutschland: Befallsstärke und Ausscheidungsintensität.

To determine the intensity of helminth infection and egg output with regard to age and sex of wild boar, stomachs and intestines of 124, lungs of 62, and livers of 39 animals from 5 enclosures in Southern Germany were examined. The evaluation of the intensity of infection showed 579.4 worms per animal for Hyostrongylus rubidus, 510.4 for Globocephalus longemucronatus, 476.9 for Oesophagostomum dentatum, 476.9 for O. quadrispinulatum, 254.2 for Metastrongylus pudendotectus, 176.9 for M. salmi, 140 for Physocephalus sexalatus, 56.9 for M. apri, 45.4 for Trichuris suis, and 3.3 for Ascaris suum. 33.6 liver flukes per infected wild boar were also found. Additionally 2 metacestodes of Taenia hydatigena on the serous liver coat of an adult sow were found. The worm burden with respect to age of the animals showed no or only minimal age related differences, whereas a large number of O. dentatum and O. quadrispinulatum was found in adults compared to younger wild boars. The evaluation of the intensity of infection with regard to sex of the animals showed higher values only for H. rubidus, O. dentatum, O. quadrispinulatum and P. sexalatus in males compared to female wild boars. The egg output intensity in the samples taken from the rectum was in average 3300 EpG for A. suum, 450.4 EpG for Oesophagostomum, 445.9 EpG for Globocephalus, 220 EpG for Trichuris, 207.9 EpG for Metastrongylus, 190.8 EpG for Hyostrongylus and 67 EpG for Capillaris. The egg output with respect to age showed higher values in younger wild boars than in adults.

[Imported infestations of nasopharyngeal parasites in dogs]. / Importierte Infestationen nasopharyngealer Parasiten beim Hund.

In this paper infestations of the nasal cavity with Myxobdella annandalei in a dog after a sojourn in Nepal and with Linguatula serrata in 2 stray dogs from Turkey, which were imported to Germany, are described.

[Helminth infections in wild boars in enclosures in southern Germany: species spectrum and infection frequency]. / Helmintheninfektionen von Wildschweinen bei Gatterhaltung in Süddeutschland: Artenspektrum und Befallshüfigkeit.

To determine the helminth fauna of wild boars, stomachs and intestines of 124, lungs of 62, and livers of 39 animals from 5 enclosures in Southern Germany were examined. The evaluation of the species composition revealed at least 13 species: Metastrongylus apri, M. salmi, M. pudendotectus, Physocephalus sexalatus, Hyostrongylus rubidus, Globocephalus longemucronatus, Ascaris suum, Trichuris suis, Oesophagostomum dentatum, O. quadrispinulatum, Fasciola hepatica, C. tenuicollis and Capillaria. The most frequently isolated species was M. salmi with 91.9%, followed by G. longemucronatus with 91.1%, M. apri with 88.7%, M. pudendotectus with 87.1%, P. sexalatus with 51.6%, O. dentatum with 46%, T. suis with 30.6%, H. rubidus with 14.5%, A. suum with 12.1% and O. quadrispinulatum with 8.1%. Differences in the prevalence of infection, determined for animals up to 1 year old, 1-2 year-old and adults, were minimal for Metastrongylus spp. and G. longemucronatus. P. sexalatus, T. suis and A. suum, however, occurred more often in 1 year old, whereas H. rubidus, O. dentatum and O. quadrispinulatum were isolated more often from adults. A correlation of the infection rate with the sex of the animals was not found.

Eimeria infections of sheep in northwest Germany.

To investigate the incidence of ovine Eimeria spp. and seasonal dynamics in oocyst output, fecal samples of sheep from three different management systems were collected monthly over a 1-year period and examined for oocysts. A total of 10 species of Eimeria were observed. The most frequent species were E. bakuensis, E. ovinoidalis, E. weybridgensis/crandallis, E. parva and E. ahsata whereas E. faurei, E. granulosa, E. intricata and E. pallida were found less often. Lambs passed larger numbers of oocysts in their feces than either ewes or yearlings.

[Parasitic diseases in sheep and goats in Germany]. / Parasitosen bei Schaf und Ziege in Deutschland.

A review is presented on the species spectrum, biology and epidemiology of endo- and ectoparasites of sheep and goats in Germany. Current therapeutic, prophylactic and metaphylactic measures are given and discussed.

[Subcutaneous dirofilariasis: infection with Dirofilaria repens]. / Subkutane Dirofilariasis: Infektion mit Dirofilaria repens.

A female patient resident in Germany is described, who had developed dirofilariasis presenting as a hard subcutaneous nodule at the glabella. Dirofilaria repens was isolated after surgical removal of the skin lesion. She was treated with diethylcarbamazine (Hetrazan) for 4 weeks. Exposures related to infection with Dirofilaria repens are discussed.

[The use of fenbendazole for controlling helminths in small zoo ruminants]. / Einsatz von Fenbendazol zur Bekämpfung von Helminthen bei kleinen Zoo-Wiederkäuern.

By means of coproscopical examination the anthelmintic efficacy of 5.0 mg fenbendazole/kg body weight and 0.714 mg/kg body weight, administered for seven consecutive days, was determined in three series of trials on small ruminants in the zoological gardens of Munich. Both doses proved to be highly effective against trichostrongylids; the egg output was reduced to an average of 100% and 80-100%, respectively. Despite of a partly lower efficiency after the multiple administration of a low level doses (7 x 0.714 mg fenbendazole/kg body weight) this is a sufficient and economical treatment for the control of the infestation with endoparasites in the - in this respect - difficult housing systems of zoological gardens.

[Endoparasitic infestation of wild hedgehogs and hedgehogs in human care with a contribution to therapy]. / Der Endoparasitenbefall wildlebender und in menschlicher Obhut befindlicher Igel mit einem Beitrag zur Therapie.

In order to confirm the prevalence of endoparasites fecal samples from 127 hedgehogs living outdoors as well as from 85 in an animal home and from 542 hedgehogs hibernating in private homes were examined. 52.0%-72.3% of the animals from natural surroundings proved to be infested with the lung worm and 72.3%-74.0% with Capillaria species of the intestine, respectively. Capillaria aerophila were found in 15.1%-40.7%, whereas coccidia (1.4%-12.9%) were less frequent. In animal homes and private care hibernating hedgehogs excreted larvae of Crenosoma striatum (23.5% and 21.0%, respectively), eggs of Capillaria species of the intestine (47.1% and 37.1%), and eggs of Capillaria aerophila (7.1% and 19.4%), but oocysts of Isospora rastegaievae were found to be predominant (44.7% and 32.3%). Proglottides of Hymenolepis erinacei and eggs of Brachylaemus erinacei appeared only in the faeces of 3 and 2 hedgehogs, respectively. Helminths of the lung and gut were already found in May, therefore it must be concluded that these parasites are able to survive the winter in the host during the hibernation period. Even young hedgehogs (400-500 g) were infected with Crenosoma and/or Capillaria spp. of the intestine, however, compared with the adults the excretion of eggs and larvae was rather low. The antiparasitic agent Ivermectin (0.3 mg/100 g body-weight) was effective against Crenosoma striatum (efficacy: 95.9%) and Capillaria spp. (100%); therefore it can be recommended as a new, well tolerated anthelmintic against nematodes of the hedgehog.