Clinical signs observed in non-indigenous and travelling dogs with vector-borne diseases
| Identified pathogen | Number of infected dogs | Number of dogs with the particular clinical sign | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Apathy | Fever | Pale mucus membranes | Icterus | Lameness/muscle pain | Gastroenteric problems | Neurological signs | Discolouration of the urine | Dermatological signs | Epistaxis | ||
| Brucella canis | 27 | 27 | 19 | 23 | 4 | 0 | 10 | 0 | 24 | 0 | 0 |
| Babesia gibsoni | 2 | 2 | 2 | 2 | 0 | 0 | 0 | 0 | 2 | 0 | 0 |
| Anaplasma phagocytophilum | 20 | 20 | 11 | 6 | 0 | 4 | 3 | 1 | 0 | 0 | 4 |
| Mycoplasma haemocanis Bartonella | 15 | 15 | 8 | 15 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| henselae | 7 | 7 | 5 | 6 | 0 | 0 | 1 | 0 | 0 | 0 | 0 |
| Ehrlichia canis | 4 | 4 | 3 | 3 | 0 | 2 | 1 | 0 | 0 | 1 | 2 |
| Hepatozoon canis Leishmania | 3 | 3 | 1 | 3 | 0 | 2 | 0 | 0 | 0 | 0 | 0 |
| infantum | 1 | 1 | 1 | 1 | 0 | 0 | 0 | 0 | 0 | 1 | 0 |
| No infection | 418 | 379 | 306 | 361 | 16 | 4 | 83 | 4 | 0 | 2 | 0 |
| Total | 497 | 458 | 356 | 420 | 20 | 12 | 98 | 5 | 26 | 4 | 6 |
| P-value | 0.002 | 0.078 | 0.016 | 0.540 | 8.517 × 10−5 | 0.999 | 0.581 | 2.200 × 10−16 | 0.121 | 1.369 × 10−5 | |
Results of vector-borne disease pathogen isolation and amplification from samples of non-indigenous and travelling dogs by country of origin or destination
| Country | Number of dogs infected with the particular pathogen | ||||||||
|---|---|---|---|---|---|---|---|---|---|
| Babesia canis | Babesia gibsoni | Anaplasma phagocytophilum | Ehrlichia cams | Hepatozoon canis | Leishmania infantum | Mycoplasma haemocanis | Bartonella henselae | Total | |
| Ukraine | 15 | 0 | 12 | 1 | 0 | 0 | 8 | 3 | 39 |
| Croatia | 6 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 7 |
| Turkey | 0 | 0 | 4 | 0 | 3 | 1 | 4 | 4 | 16 |
| Italy | 1 | 0 | 2 | 2 | 0 | 0 | 1 | 0 | 6 |
| Bulgaria | 2 | 0 | 2 | 0 | 0 | 0 | 0 | 0 | 4 |
| Albania | 1 | 1 | 0 | 0 | 0 | 0 | 2 | 0 | 4 |
| Romania | 2 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 3 |
| Total | 27 | 2 | 20 | 4 | 3 | 1 | 15 | 7 | 79 |
Primers and PCR conditions for detection and identification of Anaplasma/Ehrlichia spp_, Babesia canis, Babesia gibsoni, Bartonella henselae, Hepatozoon canis, Mycoplasma haemocanis and Leishmania infantum in samples of non-indigenous and travelling dogs
| Pathogen | Primers | Target gene | Amplicon size (base pairs (bp)) | PCR conditions | Reference |
|---|---|---|---|---|---|
| Anaplasma/ Ehrlichia spp. | EHR 521: (5′-TGT AGG CGG TTC GGT AAG TTA AAG-3′) | 16S RNA | 247 bp | 35 cycles: denaturation at | Pancholi et al. (37) |
| Babesia canis | BAB GF2: (5′-GTC TTG TAA TTG GAA TGA TGG-3′) | 18S RNA | 559 bp | 50 cycles: denaturation at | Adaszek and Winiarczyk (3) |
| Babesia gibsoni | d3: (5′-TCC GTT CCC ACA ACA-CCA GC- 3′) | P18/BgTRAP | 182 bp | 50 cycles: denaturation at | Fukumoto et al. (16) |
| Bartonella henselae | BART-LC-GEN-F: (5′-ATG GGT TTT GGT CAT CGA GT-3′) | Citrate synthase | 250 bp | 40 cycles: denaturation at | Staggemeier et al. (43) |
| Hepatozoon canis | HepF: (5′-ATA-CAT-GAG-CAA-AAT-CTC- AAC-3′) | 18S RNA | 666 bp | 34 cycles: denaturation at | Inokuma et al. (24) |
| Mycoplasma haemocanis | SYBR_TGA ACG ATG AA-3′) | 16S RNA | 103 bp | 40 cycles: denaturation at | Willi et al. (52) |
| Leishmania infantum | N13A(5′-AAC TTT TCT GGT CCT CCG GG-3′) | kinetoplast DNA minicircle | 120 bp | 40 cycles: denaturation at | Francino et al. (15) |
The effectiveness of ectoparasite prevention and probability of infection in the group of protected non-indigenous and travelling dogs
| Country | Number of protected and infected dogs | Number of non-protected and infected dogs | Number of protected and non-infected dogs | Number of non-protected and non-infected dogs | P-value | 95 % confidence interval | Odds ratio |
|---|---|---|---|---|---|---|---|
| Ukraine | 4 | 35 | 32 | 113 | 0.115 | 0.097–1.260 | 0.405 |
| Croatia | 2 | 5 | 70 | 19 | 0.009 | 0.009–0.747 | 0.111 |
| Turkey | 3 | 13 | 62 | 1 | 2.845 × 10−11 | 9.562 × 10−5–4.478 × 10−2 | 0.005 |
| Italy | 1 | 5 | 41 | 1 | 2.062 × 10−5 | 1.123 × 10−4–1.262 × 10−1 | 0.007 |
| Bulgaria,Romania and Albania | 1 | 10 | 61 | 18 | 2.007 × 10−5 | 6.752 × 10−4–2.415 × 10−1 | 0.031 |
| Total | 11 | 68 | 266 | 152 | 2.200 × 10−16 | 0.042–0.184 | 0.092 |
Haematological disorders observed in non-indigenous and travelling dogs with vector-borne diseases
| Identified pathogen | Number of infected dogs | Number of dogs with the particular haematologicaldisorder | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Anaemia | Thrombocytopaenia | Normal PLT | Leukocytosis | Leukopaenia | Normal WBC | ||||||
| Brucella canis | 27 | 27 | 27 | 0 | 2 | 21 | 4 | ||||
| Babesia gibsoni | 2 | 2 | 2 | 0 | 1 | 1 | 0 | ||||
| Anaplasma phagocytophilum | 20 | 20 | 20 | 0 | 3 | 1 | 16 | ||||
| Mycoplasma haemocanis | 15 | 15 | 9 | 6 | 10 | 0 | 5 | ||||
| Bartonella henselae | 7 | 7 | 7 | 0 | 2 | 0 | 5 | ||||
| Ehrlichia canis | 4 | 4 | 4 | 0 | 0 | 1 | 3 | ||||
| Hepatozoon canis | 3 | 3 | 1 | 2 | 3 | 0 | 0 | ||||
| Leishmania infantum | 1 | 1 | 0 | 1 | 1 | 0 | 0 | ||||
| No infection | 418 | 418 | 343 | 75 | 26 | 19 | 373 | ||||
| Total | 497 | 497 | 413 | 84 | 48 | 43 | 406 | ||||
| P-value | - | 0.191 | 0.191 | 1.807 × 10−7 | 1.949 × 10−10 | 2.200 × 10−16 | |||||
Non-indigenous and travelling dogs screened for vector-borne diseases, by country of origin or destination
| Country | Number of examined dogs | Number of dogs protected against ectoparasites | Number of infected dogs | Number of infected dogs protected against ectoparasites |
|---|---|---|---|---|
| Ukraine | 184 | 36 | 39 | 4 |
| Croatia | 96 | 72 | 7 | 2 |
| Turkey | 79 | 65 | 16 | 3 |
| Italy | 48 | 42 | 6 | 1 |
| Bulgaria | 42 | 30 | 4 | 1 |
| Albania | 36 | 31 | 4 | 0 |
| Romania | 12 | 8 | 3 | 0 |
| Total | 497 | 284 | 79 | 11 |