Serological and molecular screening for zoonotic pathogens among wild hedgehogs (Erinaceus europaeus) from urban areas of Poland

Results of hedgehog spleen sample testing in PCR, indicating the number of positive individuals and prevalence of Anaplasma spp_ and Rickettsia spp_ among the population subgroups

Variable		Total	Anaplasma spp.				Rickettsia spp.
			+	Prevalence (%)	95% CIa	P-value	+	Prevalence (%)	95% CIa	P-value
Health status	Polytrauma	15	10	66.67	41.71–84.82	0.046	4	26.67	10.90–51.95	0.325
	Respiratory disease	2	0	0.00	0.00–65.76		0	0.00	0.00–65.76
	Gastrointestinal disease	2	1	50.00	9.45–90.55		0	0.00	0.00–65.76
	Others (cachexia)	7	7	100	64.57–100		0	0.00	0.00–35.43
Sex	Male	13	8	61.64	35.52–82.29	0.671	3	23.08	8.18–50.26	0.587
	Female	13	10	76.92	49.74–91.82		1	7.69	1.37–33.31
Body weight (g)	<400	7	5	71.43	35.89–91.78	0.980	0	0.0	0.00–35.43	0.399
	400–650	12	8	66.67	39.06–86.19		3	25.00	8.89–53.23
	651–900	3	2	66.67	20.77–93.85		0	0,00	0.00–56.15
	>900	4	3	75.00	30.06–94.44		1	25.00	4.56–69.94
Total		26	18	69.23	50.01–83.50		4	15.38	6.15–33.53

The structure of the hedgehog population tested for seroprevalence

Variable		Number of individuals
Health status	Polytrauma	29
	Respiratory disease	1
	Gastrointestinal disease	1
	Others (cachexia)	4
	Healthy	4
	No data	4
Sex	Male	27
	Female	16
Body weight (g)	<400	10
	400–650	16
	651–900	13
	>900	4
Total		43

Primers and PCR conditions for detection and identification of Anaplasma/Ehrlichia spp_, Babesia spp_, Borrelia burgdorferi and Leptospira spp_ in spleen tissue samples from hedgehogs

Pathogen	Primers	Target gene	Amplicon size (base pairs)	PCR conditions	Reference
Ehrlichia spp/Anaplasma spp.	1: 5′-CTG GGG ACT ACG GTC GCA AGA C-3′2: 5′-CTC CAG TTT ATC ACT GGA AGT T-3′	16S–23S RNA	299	×1: den. at 95°C for 600 s×45: den. at 95°C for 60 s, ann. at 60°C for 60 s, ext. at 72°C for 30 s	6
Rickettsia spp.	120-2113: 5′-CGATGCTAACGTAGGTTCTT-3′120AA-2235: 5′-CCGGCTATACCGCCTGTAG-3′	rOmpB	773	×1: den. at 95°C for 180 s×40: den. at 95°C for 30 s, ann. at 50°C for 30 s, ext. at 68°C for 90 s×1: 68° for 420 s	30
Borrelia burgdorferi sensu lato	M1: 5′-ACG ATG CAC ACT TGG TGT TAA-3′M2: 5′-TCC GAC TTA TCA CCG GCA GTC A-3′	16S RNA	357	×1: den. at 85°C for 600 s×30: den. at 85°C for 30 s, ann. at 50°C for 30 s, ext. at 65°C for 60 s×1: ext. at 65°C for 600 s	20
Leptospira spp.	G1: 5′-CTG AAT CGC TGT ATA AAA GT-3′G2: 5′-GGA AAA CAA ATG GTC GGA AG-3′	16S RNA	298–320	×1: den. at 94°C for 180 s×28: den. at 94°C for 30 s, ann. at 51°C for 60 s, ext. at 72°C for 180 s×1: ext. at 72°C for 300 s	25