Vector-borne diseases imported to Poland between 2021 and 2023

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′) EHR 747: (5′-GCA CTC ATC GTT TAC AGC GTG-3′)	16S RNA	247 bp	35 cycles: denaturation at 94°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 45 s	Pancholi et al. (37)
Babesia canis	BAB GF2: (5′-GTC TTG TAA TTG GAA TGA TGG-3′)BAB GR2: (5′-CCA AAG ACT TTG ATT TCT CTC-3′)	18S RNA	559 bp	50 cycles: denaturation at 92°C for 60 s, annealing at 52°C for 60 s extension at72°C for 90 s	Adaszek and Winiarczyk (3)
Babesia gibsoni	d3: (5′-TCC GTT CCC ACA ACA-CCA GC- 3′)d4: (5′-TCC TCC TCA TCA TCC TCA TTC G-3′)	P18/BgTRAP	182 bp	50 cycles: denaturation at92°C for 60 s, annealing a52°C for 60 s extension at72°C for 90 s	Fukumoto et al. (16)
Bartonella henselae	BART-LC-GEN-F: (5′-ATG GGT TTT GGT CAT CGA GT-3′)BART-LC-HEN-R: (5′-AA ATCGACATTAGGGTAAAGTTTTT-3′)	Citrate synthase	250 bp	40 cycles: denaturation at96°C for 60 s, annealing at60°C for 60 s extension at72°C for 90 s	Staggemeier et al. (43)
Hepatozoon canis	HepF: (5′-ATA-CAT-GAG-CAA-AAT-CTC- AAC-3′)HepR (5′-CTT-ATT-ATT-CCA-TGC-TGC- AG-3′)	18S RNA	666 bp	34 cycles: denaturation at95°C for 30 s, annealing at53°C for 30 s extension at72°C for 90 s	Inokuma et al. (24)
Mycoplasma haemocanis	SYBR_TGA ACG ATG AA-3′)SYBR_Reverse 1: (5′-TGG CAC ATA GTT TGC TGT CAC TT – 3′)SYBR_Reverse 2: (5′-GCT GGC ACA TAG TTA GCT GTC ACT-3′)	16S RNA	103 bp	40 cycles: denaturation at95°C for 30 s, annealing at60°C for 60 s extension at72°C for 90 s	Willi et al. (52)
Leishmania infantum	N13A(5′-AAC TTT TCT GGT CCT CCG GG-3′)N13B (5′-CCC CCA GTT TCC CGC CC-3′)	kinetoplast DNA minicircle	120 bp	40 cycles: denaturation at94°C for 30 s, annealing at58°C for 30 s extension at72°C for 30 s	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