BackgroundAngiostrongylus vasorum is a potentially fatal canine nematode. Due to the high variability of clinical signs and the often chronic and subtle course of the infections, the diagnosis is particularly challenging. A rapid in-clinic assay (Angio Detect™ Test, IDEXX Laboratories, Westbrook, Maine, USA) for the serological detection of circulating antigen and intended for routine in-clinic diagnosis has been evaluated.MethodsSensitivity was calculated with sera from 39 naturally infected dogs confirmed by Baermann-Wetzel analysis, while sera of 38 experimentally infected dogs were used for follow-up analyses, of which 10 were treated with imidacloprid/moxidectin. Cross-reactivity was tested with a total of 123 samples from dogs with proven parasitic infections with Toxocara canis (n = 21), Ancylostoma caninum (n = 4), Crenosoma vulpis (n = 18), Oslerus osleri (n = 3), Eucoleus aerophilus, (n = 6), Dirofilaria immitis (n = 28), Dirofilaria repens (n = 20), Acantocheilonema reconditum (n = 10) or Dipetalonema dracunculoides (n = 10) or multiple infections (n = 3). All sera were tested with the Angio Detect™ Test and with an ELISA for detection of circulating antigen of A. vasorum.ResultsThe sensitivity of the Angio Detect™ Test was 84.6% (95% C.I. 69.5 - 94.1%), while specificity was 100% (95% C.I. 97.6 - 100%). The sensitivity of the ELISA (94.9%, 95% C.I. 82.7 – 99.3%) was comparable with previous evaluations. In experimentally infected dogs, earliest positive results with the Angio Detect™ Test were observed 9 weeks post inoculation and 5 weeks later all sera were Angio Detect™ Test positive. After anthelmintic treatment, seropositive dogs turned negative again within 3 to 7 weeks after treatment. The evaluation of the colour intensity of the test strips confirmed the delay of approximately 3-4 weeks for antigen detection by the Angio Detect™ Test compared to the ELISA and its correlation with the time after infection.ConclusionsThis study provided evidence of a good sensitivity and a very high specificity of the rapid device Angio Detect™ Test for detection of circulating A. vasorum antigen in dogs with suspected canine angiostrongylosis, representing a very simple and useful tool to be broadly applied in veterinary practices. The rapid detection of infected dogs is a key point for initiating an indispensable and urgent therapy.

BackgroundConsidering the increasing importance of small animals travel medicine and the spread of filariae with zoonotic potential to non-endemic European areas, routine filarial diagnosis in dogs is becoming important. Dirofilaria immitis, D. repens, Acanthocheilonema dracunculoides and A. reconditum are the most common canine filarial nematodes presenting blood circulating microfilariae (mf) which can be differentiated to species level by the acid phosphatase activity patterns or by PCR. Available data on the size of the mf vary considerably in the literature. The aim of this study was to validate morphometric criteria for filarial identification in blood samples of dogs after concentration of mf with the modified Knott’s technique.MethodsMorphometric analysis of 10 mf from samples identified to species level by acid phosphatase activity and partially confirmed by PCR were performed with specimens from 377 dogs.ResultsThe mean length and width of D. immitis mf from 60 dogs were 301.77±6.29 μm and 6.30±0.26 μm, of D. repens mf from 171 dogs 369.44±10.76 μm 8.87±0.58 μm, of A. dracunculoides mf from 133 dogs 259.43±6.69 μm and 5.09±0.47 μm and of A. reconditum mf from 13 dogs 264.83±5.47 μm and 4.63±0.52 μm.For a subset of 30 samples, morphometric analysis was repeated with identical results in two laboratories. Furthermore, the size of mf concentrated and fixed by the Knott’s technique was shown to be stable over 105 days.ConclusionsThe Knott’s test enables to clearly distinguish between D. immitis, D. repens and Acanthocheilonema spp. However, due to the overlapping size ranges of A. dracunculoides and A. reconditum, biochemical or molecular methods are required to distinguish these two species.

BackgroundWith the import of pets and pets taken abroad, arthropod-borne diseases have increased in frequency in German veterinary practices. This is reflected by 4,681 dogs that have been either travelled to or relocated from endemic areas to Germany. The case history of these dogs and the laboratory findings have been compared with samples collected from 331 dogs living in an endemic area in Portugal. The various pathogens and the seroprevalences were examined to determine the occurrence of, and thus infection risk, for vector-borne pathogens in popular travel destinations.Results4,681 dogs were examined serological for Leishmania infantum, Babesia canis and Ehrlichia canis. Buffy coats were detected for Hepatozoon canis and blood samples were examined for microfilariae via the Knott's test. The samples were sent in from animal welfare organizations or private persons via veterinary clinics. Upon individual requests, dogs were additionally examined serological for Anaplasma phagocytophilum, Borrelia burgdorferi and Rickettsia conorii. Overall B. canis was the most prevalent pathogen detected by antibody titers (23.4%), followed by L. infantum (12.2%) and E. canis (10.1%). Microfilariae were detected in 7.7% and H. canis in 2.7% of the examined dogs. In 332/1862 dogs A. phagocytophilum, in 64/212 B. burgdorferi and in 20/58 R. conorii was detected. Of the 4,681 dogs, in total 4,226 were imported to Germany from endemic areas. Eighty seven dogs joined their owners for a vacation abroad. In comparison to the laboratory data from Germany, we examined 331 dogs from Portugal. The prevalence of antibodies/pathogens we detected was: 62.8% to R. conorii, 58% to B. canis, 30.5% to A. phagocytophilum, 24.8% to E. canis, 21.1% to H. canis (via PCR), 9.1% to L. infantum and 5.3% to microfilariae.ConclusionsThe examination of 4,681 dogs living in Germany showed pathogens like L. infantum that are non-endemic in Germany. Furthermore, the German data are similar in terms of multiple pathogen infection to the data recorded for dogs from Portugal. Based on these findings the importation of dogs from endemic predominantly Mediterranean regions to Germany as well as travelling with dogs to these regions carries a significant risk of acquiring an infection. Thus we would conclude that pet owners seek advice of the veterinarians prior to importing a dog from an endemic area or travel to such areas. In general, it might be advisable to have a European recording system for translocation of dogs.

This study has provided evidence for the natural occurrence of sandflies in Germany. Two species belonging to the genus Phlebotomus were detected. Phlebotomus (Larroussius) perniciosus, a proven vector of leishmaniasis, was found in association with an autochthonous case of canine leishmaniasis near Kaiserslautern. Two hundred thirty-seven specimens of Phlebotomus (Transphlebotomus) mascittii were caught in 17 different locations in Baden-Wuerttemberg and Rhineland-Palatinate. The northernmost finding in Germany (and Europe) was near Cochem (Moselle). P. mascittii has not yet been confirmed as a vector of leishmaniasis, but its competence is strongly suspected. In addition to the detection of the vector, since 1991, there have been 11 cases of leishmaniasis in Germany in which an autochthonous origin was confirmed or which were highly likely to have been of indigenous origin. Data from the German meteorological service indicate that Germany currently has a Mediterranean climate, with an annual average temperature of 10 degrees C being reached or exceeded in several regions. This type of climate is also appropriate for the living conditions of sandflies. Therefore, it is assumed that sandflies have a greater geographical distribution in Germany than the first studies suggested, being mainly restricted to the southern region of Baden-Wuerttemberg. The risk of an autochthonous canine infection occurring in Germany is very low. The rapidly increasing number of imported cases of leishmaniasis in dogs means that special attention must be focused on veterinary advice to dog owners about prophylaxis. The results indicate that the use of repellents and preventive behavioural measures is vital.

BackgroundCanine leishmaniosis (CanL) is a zoonotic disease caused by Leishmania (L.) infantum. It is endemic to several tropical and subtropical countries but also to the Mediterranean region. It is transmitted by phlebotomine sandflies but occasional non-vector transmissions have been reported, including vertical and horizontal transmission.FindingsThe authors report a case of CanL in a female boxer dog from Dusseldorf, Germany, that had never been in an endemic region. A serum sample from the bitch was tested positive for antibodies against Leishmania (IFAT 1:2,000, ELISA 72). The bitch had whelped three litters, and one puppy from the third litter was also found to be seropositive for Leishmania antibodies (IFAT 1:4,000, ELISA 78).ConclusionsUp to now, despite intensive searching, the occurrence of sandflies could not be proved in the bitch's region of origin. Thus, vertical and horizontal transmission are to be discussed as possible ways of infection. This may be the first report of venereal and vertical transmission of L. infantum in naturally infected dogs in Germany.