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DOI: 10.

2478/s11686-014-0289-0
W. Stefaski Institute of Parasitology, PAS
Acta Parasitologica, 2014, 59(4), 661665; ISSN 1230-2821

The first clinically manifested case of angiostrongylosis


in a dog in Slovakia
Martina Miterpkov1, Zuzana Hurnkov1, 2 and Artur P. Zalewski3
1
Institute of Parasitology of the Slovak Academy of Sciences, Hlinkova 3, 040 01 Koice, Slovakia,
2
University of Veterinary Medicine and Pharmacy, Komenskho 73, 040 01 Koice, Slovakia,
3
Bayer Sp. z o.o. Animal Health, Al. Jerozolimskie 158, 02-326 Warszawa, Poland

Abstract
The first clinical case of canine angiostrongylosis from Slovakia, previously infection-free country, is described. 18-month old
male Bernese mountain dog living in south-eastern part of Slovakia showed poor health condition characterized by weight loss,
irritating cough, dispnoe, intense salivation, vomiting and bilateral scleral bleeding. Two times even the acute physical collapse
occurred. Blood analysis was provided and revealed increase of total protein, eosinophilia, monocytosis, and mild thrombocy-
topenia. Anaemia characterized by reduced number of erythrocytes and reduced levels of haemoglobin, packed cell volume and
iron was also diagnosed. Larvoscopic Baermann technique revealed the presence of Angiostrongylus first stage larvae. Infected
dog excreted larvae in high numbers in 10 g of the faecal material more than 800 larvae were counted. DNA analysis using
PCR confirmed the presence of Angiostrongylus vasorum species. The first clinical case of angiostrongylosis has evidenced that
the new life-threatening parasitic disease of dogs has spread to the territory of Slovakia. A serious effort is therefore inevitable
to increase the professional awareness and knowledge on diagnosis, treatment and prevention.

Keywords
Angiostrongylus vasorum, dog, clinical signs, Slovakia

Introduction The first autochthonous case of canine angiostrongylosis


in Slovakia was diagnosed in 7-month old Maltese pinch liv-
The present study describes the first clinical report of an- ing in Kosice (eastern Slovakia). The dog showed no clinical
giostrongylosis in a naturally infected dog from Slovakia. signs of infection (Hurnkov et al. 2013). After treatment,
The agent of infection, Angiostrongylus vasorum, is the consecutive follow-up examinations were negative for the
metastrongylid nematode with indirect life cycle. Definitive presence of A. vasorum larvae in faeces.
hosts are carnivores and several slug species and frogs play
the role of the intermediate host (Koch and Willesen 2009). Case report
The parasite, known also as the French worm, causes severe,
often fatal cardiopulmonal disease of dogs. Herein we present the first clinically manifested case of ca-
In recent years, the awareness of veterinarians and breed- nine angiostrongylosis in Slovakia, diagnosed in an 18-month
ers on this parasitic disease has been growing due to reason- old male Bernese mountain dog. The dog was found strayed in
able spread of A. vasorum outside its endemic areas of the village Horovce (Michalovce district, Eastern Slovak Low-
southeastern France, England, Denmark and Canada (Boltet land) in June 2013. Estimated age of the individual at that time
et al. 1994). The parasite has recently appeared in European was approx. 9 months. The dog was rescued through the shel-
countries where it previously did not exist in central Europe ter and has been living with new owners in Kosice, eastern
A. vasorum was recorded in Hungary (Majoros et al. 2010) Slovakia.
and a serological study confirmed its presence in Poland At the time of adoption, the dog showed poor general con-
(Schnyder et al. 2013). In 2013 the infection was confirmed dition with sporadic cough and was severely infested with
during routine faecal examination in a dog in the Czech Re- ticks. Finding of routine faecal examination performed at vet-
public (Svobodov, personal communication) and in Slovakia erinary clinic stated the presence of Trichuris spp. eggs,
(Hurnkov et al. 2013). strongylid eggs (Ancylostoma/Uncinaria spp.), and some un-

*Corresponding author: miterpak@saske.sk


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662 Martina Miterpkov et al.

specified larvae. The dog was given fenbendazole (Fenbion, the dorsal side and also small indentation on the ventral side.
250 tbl. a.u.v., Mevak) in two doses and subsequently his Larvae isolated from fresh faeces were all coiled. Infected dog
overall health improved. The owner observed persisting mod- excreted larvae in high numbers in 10 g of the faecal mate-
erate intermittent cough, primarily after physical effort. Blood rial more than 800 larvae were counted. Considering the stray
sample was therefore examined for dirofilariosis with nega- episode in the case-history, it is possible that the dog was com-
tive result. pelled to feed on alternative food sources, including snails and
After four months the health condition of the dog deterio- thus we can assume that the infection pressure was eminent
rated rapidly and significant lethargy, loss of appetite and in this case.
weight loss of 14 kg appeared, associated with strenuous walk- DNA from larvae (10 specimens) was extracted using the
ing and hunched posture. Two times even the acute physical DNeasyBlood&Tissue Kit (QIAGEN Group, Germany).
collapse occurred. Hemorrhagic diarrhoea, intense salivation, PCR reaction for the amplification of rDNA second internal
vomiting, difficulties with urination and haematuria were also transcribed spacer (ITS2) was performed in a final volume of
reported. Scleral bleeding was evident bilaterally. The dog suf- 25 l using specific A. vasorum pairs of primers AV5 and AV4
fered from severe irritating cough and dyspnoea. designed by Al-Sabi et al. (2010). Template DNA (5 l) was
With regard to the anamnesis, the faecal sample from the amplified by PCR reaction started with a pre-heating step at
dog was delivered to the Institute of Parasitology SAS, where 94C for 2 min and 38 cycles consisted of denaturing at 94C
the modified larvoscopic Baermann technique (Willesen et al. for 30 s, annealing at 57C for 30 s and extension at 72C for
2004) was performed and revealed the presence of morpho- 30 s. The PCR was terminated with a final extension at 72C
logically distinctive Angiostrongylus vasorum first stage lar- for 7 min. Amplicons were visualized on a 1.5% agarose gel.
vae (Fig. 1.). The length of the larvae ranged between 320 and DNA analysis confirmed the presence of A. vasorum in in-
360 m and their caudal end showed distinct indentation on fected dog.

Fig 1. The first stage larvae (L1) of Angiostrongylus vasorum, immobilised


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Canine angiostrongylosis in Slovakia 663

Table I. Clinical and abnormal laboratory findings registered at diagnosis in Angiostrongylus vasorum
in naturally infected dog

Clinical findings
poor general conditions, lethargy, appetite and weight loss,
cough, dispnoe, fever, acute collapse,
hemorrhagic diarrhoea, vomitus, intense salivation, haematuria,
bilateral scleral bleeding
Abnormal laboratory findings
Parameter (reference range) Haematological findings
Total Protein (5575 g/l) 95 g/l
Thrombocytes (200500 109/l) 145109/l
PCV (0.370.55 l/l) 0.29 l/l
Haemoglobin (120180 g/l) 101 g/l
Erythrocytes (5.58.51012/l) 4.11012/l
Iron (1621 mol/l) 13.7 mol/l
Eosinophils (05%) 12%
Monocytes (210%) 23%

The blood sample of the dog was also subjected to labora- of haemoglobin, packed cell volume (PCV), and iron was also
tory tests and haematological and biochemical profile was pro- diagnosed (Table I.). Due to several specific clinical signs, the
vided. Blood analysis revealed several abnormal findings dogs blood was also examined for the presence of Babesia
(Table I.). canis with negative result.
The increase of total protein, eosinophilia, monocytosis, Immediately after diagnosis the dog was treated with two
and mild thrombocytopenia were recorded. Anaemia charac- administrations of spot-on imidacloprid/moxidectin solution
terized by reduced number of erythrocytes and reduced levels (Advocate, Bayer Animal Health). One month later, the fae-

Fig. 2. Location of two cases of canine Angiostrongylus vasorum infection in eastern Slovakia (AUT Austria, CZE Czech
Republic, HUN Hungary, POL Poland, UKR Ukraine)
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664 Martina Miterpkov et al.

cal Baermann examination was negative, respiratory signs dis- angiostrongylosis confirms the fenbendazole administration
appeared and general health condition has improved quickly. being highly effective, when using proved therapeutic dose
2050 mg/kg for 521 days (Chapman et al. 2004). In some
cases even the prolonged use of the preparation for a period of
Discussion and Conclusions 35 days is necessary (Gallagher et al. 2012). Therapeutically
effective is also the administration of imidacloprid/moxidectin
The first diagnosed case of canine angiostrongylosis in Slo- spot-on solution in single topical dose (Willesen et al. 2007),
vakia published in 2013 (Hurnkov et al. 2013) already which was approved in the treatment of our patient.
pointed out that local climate and overall ecological condi- The first clinical case of angiostrongylosis has evidenced
tions comply with the requirements of the parasites develop- that the new life-threatening parasitic disease of dogs has
ment and circulation. Herein described second case, spread to the territory of Slovakia. A serious effort is therefore
manifested by significant clinical symptoms and almost fatal inevitable to increase the professional awareness and knowl-
course, has clearly confirmed this assumption. edge on diagnosis, treatment and prevention. Our experience
Hitherto, both reported angiostrongylosis cases in Slova- (particularly with dirofilariosis prevention and control) indi-
kia have been diagnosed in dogs from the region of south-east- cates that the monitoring and research related to emerging dis-
ern Slovakia (Fig. 2.). Here presented second case was eases is very challenging (Miterpkov et al. 2012), and longer
ascertained in an individual coming from the area of the East- time is required for establishing effective and continuous co-
ern Slovak Lowland characterized by specific ecological con- operation with veterinary practitioners.
ditions. Major part of the territory is covered by woodland
meadows, flood plain forests and swamps, the winters are very
mild, often without snow cover, and the average temperature Acknowledgements. The work was supported by the science grant
is among the highest in Slovakia. This geographic and climate agency VEGA (grant no. 2/0011/12).
terms provide suitable conditions for both, survival and re-
production of many parasite vectors from mosquitoes and
ticks to snails. Recent parasitological and epidemiological References
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Received: March 13, 2014


Revised: June 2, 2014
Accepted for publication: July 29, 2014

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