Martin Knaus

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.

Release of Lungworm Larvae from Snails in the Environment: Potential for Alternative Transmission Pathways

Background Gastropod-borne parasites may cause debilitating clinical conditions in animals and humans following the consumption of infected intermediate or paratenic hosts. However, the ingestion of fresh vegetables contaminated by snail mucus and/or water has also been proposed as a source of the infection for some zoonotic metastrongyloids (e.g., Angiostrongylus cantonensis). In the meantime, the feline lungworms Aelurostrongylus abstrusus and Troglostrongylus brevior are increasingly spreading among cat populations, along with their gastropod intermediate hosts. The aim of this study was to assess the potential of alternative transmission pathways for A. abstrusus and T. brevior L3 via the mucus of infected Helix aspersa snails and the water where gastropods died. In addition, the histological examination of snail specimens provided information on the larval localization and inflammatory reactions in the intermediate host. Methodology/Principal Findings Twenty-four specimens of H. aspersa received ~500 L1 of A. abstrusus and T. brevior, and were assigned to six study groups. Snails were subjected to different mechanical and chemical stimuli throughout 20 days in order to elicit the production of mucus. At the end of the study, gastropods were submerged in tap water and the sediment was observed for lungworm larvae for three consecutive days. Finally, snails were artificially digested and recovered larvae were counted and morphologically and molecularly identified. The anatomical localization of A. abstrusus and T. brevior larvae within snail tissues was investigated by histology. L3 were detected in the snail mucus (i.e., 37 A. abstrusus and 19 T. brevior) and in the sediment of submerged specimens (172 A. abstrusus and 39 T. brevior). Following the artificial digestion of H. aspersa snails, a mean number of 127.8 A. abstrusus and 60.3 T. brevior larvae were recovered. The number of snail sections positive for A. abstrusus was higher than those for T. brevior. Conclusions Results of this study indicate that A. abstrusus and T. brevior infective L3 are shed in the mucus of H. aspersa or in water where infected gastropods had died submerged. Both elimination pathways may represent alternative route(s) of environmental contamination and source of the infection for these nematodes under field conditions and may significantly affect the epidemiology of feline lungworms. Considering that snails may act as intermediate hosts for other metastrongyloid species, the environmental contamination by mucus-released larvae is discussed in a broader context.

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.

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.

Lungworms and gastrointestinal parasites of domestic cats: a European perspective

With the exception of Aelurostrongylus abstrusus, feline lungworms have been poorly studied. Information on their distribution is patchy and mostly limited to case reports. In this study, the occurrence of feline lungworms and co-infecting gastrointestinal parasites has been investigated in 12 European countries (i.e. Austria, Belgium, Bulgaria, France, Greece, Hungary, Italy, Portugal, Romania, Spain, Switzerland and the United Kingdom). An average of 10 domestic cats, with regular outdoor access, was sampled each month for 12months, and freshly passed faeces were collected. Stools were processed using a McMaster assay and a quantitative Baermann-Wetzel method. Animals positive for lungworms and/or gastrointestinal parasites were treated with a formulation containing fipronil, (S)-methoprene, eprinomectin, and praziquantel (Broadline®, Merial), and re-sampled 28days post-treatment. The association between lungworm infection and risk factors was analysed using statistical medians/means and the efficacy of the treatment against each lungworm species was assessed. Of 1990 cats sampled, 613 (30.8%) were positive for at least one parasite, while 210 (10.6%) were infected by lungworms. The prevalence of lungworm infection varied between the sampled sites, with the highest recorded in Bulgaria (35.8%) and the lowest in Switzerland (0.8%). None of the cats from Austria or the United Kingdom were infected by lungworms. Aelurostrongylus abstrusus was the species most frequently detected (78.1%), followed by Troglostrongylus brevior (19.5%), Eucoleus aerophilus (14.8%) and Oslerus rostratus (3.8%). The overall efficacy of the treatment was 99% for A. abstrusus and 100% for T. brevior, O. rostratus and E. aerophilus. Data presented provide a comprehensive account of the diagnosis, epidemiology and treatment of feline lungworms in Europe, as well as of the occurrence of co-infections by gastrointestinal parasites.

Efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel against larval and adult stages of the cat lungworm, Aelurostrongylus abstrusus

The efficacy of a novel topical combination of fipronil 8.3% w/v, (S)-methoprene 10% w/v, eprinomectin 0.4% w/v, and praziquantel 8.3% w/v (BROADLINE(®),(1) Merial) against larval and adult Aelurostrongylus abstrusus lungworms in cats was assessed in a controlled laboratory study. The study included 48 purpose-bred, short-haired cats which were each inoculated with 225 infective A. abstrusus larvae. The cats were formed into eight blocks based on pre-treatment bodyweight and were then, within each block, randomly allocated to one of six treatment groups: untreated control; treated once when A. abstrusus were expected to be third-stage larvae (4 days post inoculation [dpi]), fourth-stage larvae (7 dpi), immature adults (14 dpi) or adult nematodes (32 dpi), or treated twice, once when A. abstrusus were expected to be third-stage larval and once again when A. abstrusus were expected to be adult nematodes (4 dpi+32 dpi). Cats weighing ≥ 0.8-2.5 kg received one 0.3 mL applicator and cats weighing >2.5-7.5 kg received one 0.9 mL applicator. For determination of the efficacy of treatments, lungworm larval counts were established on faecal samples collected from all cats 32, 39, 46, 53 and 60 dpi. At each occasion from 46 dpi on, cats treated with fipronil, (S)-methoprene, eprinomectin and praziquantel had significantly lower A. abstrusus larval counts than the untreated controls with percentage reductions of 91.6% (cats treated 14dpi; P=0.012), ≥ 98.9% (cats treated either 4 dpi, 7 dpi or 32 dpi; P<0.001) or >99.9% (cats treated 4 dpi+32 dpi; P<0.001) at 60 dpi. Thus, the novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel was highly effective in the prevention and treatment of A. abstrusus lungworm infection in cats.

Rickettsia felis and Bartonella spp. in fleas from cats in Albania.

Fleas can serve as vectors for bacterial pathogens like Bartonella and Rickettsia species, which have been isolated worldwide. However, the knowledge of the epidemiology of vector-borne diseases in general and thus on flea-borne diseases in Albania is limited. Therefore, from 78 free-roaming cats in Tirana, Albania, fleas (371 Ctenocephalides felis and 5 Ctenocephalides canis) were collected to examine them for the presence of Rickettsia and Bartonella species. Ten of the 371 C. felis (2.7%) were positive for Rickettsia felis, and 24 (6.5%) for Bartonella spp. (B. henselae and B. clarridgeiae). In total, fleas from 15 cats (19.2%) were positive for either one or the other of the pathogens. The results of this study provided evidence for the presence of R. felis (causing flea-borne spotted fever) and Bartonella spp. (causing cat scratch disease) in Albania. Thus, these infectious diseases should be considered as differential diagnoses when febrile symptoms are presented, especially after contact with cats or their fleas.

Efficacy of Broadline® spot-on against Aelurostrongylus abstrusus and Troglostrongylus brevior lungworms in naturally infected cats from Italy

The increasing reports of Aelurostrongylus abstrusus infection and the new information on Troglostrongylus brevior have spurred the interest of the scientific community towards the research of pharmaceutical compounds effective against both pathogens. A novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel (Broadline®, Merial) has been released for the treatment of a variety of feline parasitic infections. The present study reports the efficacy of this spot-on in treating cats naturally infected by feline lungworms. Client owned cats (n=191) were enrolled from three geographical areas of Italy and faecal samples were examined by floatation and Baermann techniques. Twenty-three individuals were positive for L1 of A. abstrusus (n=18) or T. brevior (n=3) or for both species (n=2) and they were topically treated with Broadline®. Seventeen of them were also concomitantly infected by other parasites. Four weeks after treatment, faecal samples were collected and examined to assess the efficacy of a single administration of the product. Based on lungworm larvae counts, the efficacy of the treatment was 90.5% or 100% for A. abstrusus or T. brevior, respectively. Cats released significantly lower amounts of lungworm larvae after treatment compared to pre-treatment (p<0.0001). All but three cats were negative for other nematodes after treatment and all cats recovered from respiratory signs. Results of this study indicate that a single administration of the topical combination fipronil, (S)-methoprene, eprinomectin and praziquantel is effective and safe for the treatment of A. abstrusus and/or T. brevior infections in cats living under field conditions.

Efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel against feline urinary bladder worm (Capillaria plica) infection.

Infection with urinary capillarid bladder worms has been observed in cats worldwide. Although considered as generally causing no or little harm, infection with urinary capillarids may be associated with clinical disease which requires an appropriate treatment including the use of anthelmintics. Therefore, the efficacy of a novel topical combination formulation of fipronil 8.3% (w/v), (S)-methoprene 10% (w/v), eprinomectin 0.4% (w/v), and praziquantel 8.3% (w/v) (BROADLINE(®), Merial) was evaluated against urinary capillarids in naturally infected cats. Sixteen European Short Hair cats (5 male, 11 female) with capillarid eggs in their urine pre-treatment were included in the study. At the time of treatment, the cats were approximately ten months to eight years old and weighed 1.6-3.6 kg. Cats were ranked based on decreasing bodyweight and then randomly allocated within replicates of two animals to one of the treatment groups. Each cat in the treated group received one topical application of the combination product at the minimum therapeutic dose of 0.12 mL/kg body weight delivering 10mg fipronil+12 mg (S)-methoprene+0.5mg eprinomectin+10mg praziquantel per kilogram of body weight while the cats allocated to the control group remained untreated. For parasite recovery, identification and count, cats were euthanized humanely 14 days after treatment. All untreated cats harboured Capillaria plica in their urinary bladders (range 4-12), while no capillarids were recovered from the eight treated cats. Thus, the efficacy of the novel topical combination against C. plica was 100%. All cats accepted the treatment well based on post-treatment observations and daily observations thereafter. No adverse events or other health problems were observed during the study.