Mixed-sex clusters on grass blades: breeding strategy of the ornate dog tick, Dermacentor reticulatus.

BACKGROUND: The ornate dog tick Dermacentor reticulatus is second only to the hard tick Ixodes ricinus in terms of importance as a vector of infectious organisms, especially of Babesia canis, the agent of canine babesiosis. Both the geographical range and local densities of D. reticulatus are steadily increasing in many regions of Europe. In the present study, we tested the hypothesis that D. reticulatus possesses an efficient breeding strategy that allows for a rapid increase in tick numbers and densities through the formation of mixed-sex clusters/aggregations while questing in the environment. METHODS: An observational study was carried out in the spring of 2023, at three sites in two regions in Central and North-Eastern Poland, both characterised by high tick densities. At each site, a 400-m-long transect was inspected for questing ticks. All noted ticks were collected, and tick numbers and sexes per stem were recorded. Differences in tick distribution by site and sex were analysed statistically. RESULTS: A total of 371 D. reticulatus (219 females, 152 males) ticks were collected from 270 grass stems over a combined 1200 m of transect. The majority of grass stems (74.4%) were occupied by just a single individual, with two-tick clusters the second most common category. The maximum number of D. reticulatus individuals observed on a single grass stem was six. Mixed-sex clusters were significantly more common than single-sex clusters at all three sites. With study sites combined, mixed-sex clusters accounted for 17.4% (95% confidence limit [95% CL] 13.9-21.6%) of observations, while for multiple males and multiple females, the values were 2.6% (95% CL: 1.4-4.7%) and 5.6% (95% CL: 3.7-8.3%), respectively. CONCLUSIONS: Mixed-sex clusters of D. reticulatus ticks were significantly more common than single-sex clusters, which we hypothesise reflects an efficient, likely pheromone-mediated breeding strategy of this expansive tick species.

The first report on Hepatozoon canis in dogs and wolves in Poland: clinical and epidemiological features.

BACKGROUND: Canine hepatozoonosis caused by Hepatozoon canis is a common infection in dogs, with frequent case reports from the Mediterranean region and more recently from several Central European countries, such as Hungary and Germany. Despite the high prevalence of H. canis in red foxes, no infections have been reported to date in dogs in Poland. We describe here the first autochthonous cases of H. canis infection in dogs, including their clinical features, and report the prevalence of H. canis in grey wolves from different regions of Poland. METHODS: Thin smears prepared from blood samples collected from dogs were evaluated by microscopic examination. A total of 60 wolves and 47 dogs were tested. Infections were confirmed by PCR and sequencing. RESULTS: Gamonts of H. canis were found in > 50% of the neutrophils of two dogs and in < 10% of the neutrophils in another five dogs. Molecular typing by PCR sequencing of the 18S ribosomal RNA gene fragment confirmed infections in 11 dogs from different regions of Poland, in 2.7% of dogs attending veterinary practices in central Poland and in 35% of wolves from various geographical regions of Poland. Clinical features manifested mostly in older dogs, and the most common signs were anaemia and apathy. Young dogs usually remained asymptomatic. CONCLUSIONS: This is the first report of H. canis infection in dogs and wolves in Poland. Although the exact vector of the parasite is not known, veterinary practitioners should be aware of this new parasitosis and should consider appropriate diagnostics to confirm/exclude this infection. Further studies are needed to understand the transmission routes of H. canis in domestic and wild canids in Poland.

Occurrence of Dirofilaria repens in wild carnivores in Poland.

Dirofilaria repens is an expanding vector-borne zoonotic parasite of canines and other carnivores. Sub-clinically infected dogs constitute the most important reservoir of the parasite and the source of infection for its mosquito vectors. However, occurrence of D. repens infection in wild animals may contribute to the transmission of the parasite to humans and may explain the endemicity of filariae in newly invaded regions. The aim of the current study was to determine the occurrence of D. repens in 511 blood and spleen samples from seven species of wild carnivores (wolves, red foxes, Eurasian badgers, raccoons, raccoon dogs, stone martens, and pine martens) from different regions of Poland by means of a PCR protocol targeting the 12S rDNA gene. Dirofilaria repens-positive hosts were identified in seven of fourteen voivodeships in four of the seven regions of Poland: Masovia, Lesser Poland, Pomerania and Warmia-Masuria. The highest prevalence was found in Masovia region (8%), coinciding with the highest previously recorded prevalence in dogs in Central Poland. The DNA of Dirofilaria was detected in 16 samples of three species (total prevalence 3.13%). A low and similar percentage of positive samples (1.9%, 4.2% and 4.8%) was recorded among badgers, red foxes, and wolves, respectively. Dirofilaria repens-positive hosts were identified in seven of fourteen voivodships. Based on detection in different voivodeships, D. repens-positive animals were recorded in four out of the seven regions of Poland: in Masovia, Lesser Poland, Pomerania, and Warmia-Masuria. The highest prevalence of filariae was found in Masovia region (8%), reflecting the highest previously recorded prevalence in dogs (12-50%) in Central Poland. In summary, we conducted the first comprehensive study on the epidemiology of D. repens in seven species of wild hosts in all seven regions of Poland and identified the first case of D. repens infection in Eurasian badgers in Poland and the second in Europe.

Haematological indices and immune response profiles in dogs naturally infected and co-infected with Dirofilaria repens and Babesia canis.

Co-infections with Dirofilaria repens and Babesia canis are rarely reported in the literature and there is very limited knowledge of their impact on canine health. Central Poland is endemic for both parasites, posing a risk of co-infections in dogs. To evaluate the impact of co-infection with B. canis and D. repens on canine health, four groups of dogs were examined: healthy dogs, dogs infected with B. canis, dogs infected with D. repens and dogs co-infected with both species. Blood parameters indicative of anaemia, kidney and liver damage were analysed statistically. Additionally, expression levels of immune response genes were quantified and compared, to define the type of immune response typically encountered in single- and co-infections. In dogs infected with D. repens, no major alterations in blood parameters were observed. Dogs infected with B. canis suffered from anaemia, kidney and liver insufficiency. In contrast, dogs co-infected with D. repens and B. canis showed milder alternation in blood biochemical parameters associated with liver (ALP activity) and kidney (serum urea and creatinine levels) dysfunction, compared to dogs infected only with B. canis. The expression of genes associated with cellular (Th1-mediated) (STAT4 and INF-γ), humoral (Th2-mediated) (STAT6, GATA3, SOCS3, IL-13) and regulatory (IL-10) responses was quantified. For this analysis, dogs infected with B. canis were divided into two groups-'Babesia 1' (mild babesiosis), 'Babesia 2' (severe babesiosis). All the tested factors, except INF-γ, were found to be expressed in dogs infected with D. repens. In 'Babesia 1' dogs, expression of GATA3 was highest, while in 'Babesia 2'-INF-γ and SOCS3 dominated. IL-13 expression was predominant in dogs infected with D. repens, and STAT6 and IL-10 predominated in dogs with co-infections.

Tick-Borne Pathogens, Babesia spp. and Borrelia burgdorferi s.l., in Sled and Companion Dogs from Central and North-Eastern Europe.

Ticks are important vectors of numerous pathogens of medical and veterinary significance. The aim of the current study was to determine the prevalence of Babesia spp. and Borrelia burgdorferi s.l. in sled and pet dogs from Central and North-Eastern Europe. Neither Babesia spp. nor Borrelia burgdorferi s.l. infections were detected in sled dogs from seven countries (Poland, Lithuania, Latvia, Estonia, Belarus, Russia and Finland). The DNA of Babesia spp. was detected in 100% of symptomatic and 5.4% of asymptomatic pet dogs from Poland. Similarly, the DNA of Babesia spp. was identified in 82% of symptomatic and 3.8% of asymptomatic pet dogs from Ukraine. The DNA of Borrelia burgdorferi s.l. was detected in 4.4% of pet dogs. Molecular typing confirmed the presence of Babesia canis and Borrelia burgdorferi sensu stricto (s.s.) in selected samples. Four dogs were co-infected by B. canis and Borrelia burgdorferi s.l. Tick-borne pathogens constitute a serious health threat to pet dogs in Central and South-Eastern Europe, but were not observed among sled dogs from the same region of Europe nor in the Baltic countries.