Microscopic and molecular analysis of Babesia canis in archived and diagnostic specimens reveal the impact of anti-parasitic treatment and postmortem changes on pathogen detection

Table 1 Parasite burden and genotyping results from archival cytological slides (Group A)

Samples	Tissue^a	Year of preparation	Parasite burden	PCR protocols^b
Samples	Tissue^a	Year of preparation	Parasite burden	cytb	1	2	3
Postmortem samples
A-1	Kidney	2003	286 (H)	+	+	+	+
A-2	Kidney	2004	527 (VH)	+	–	+	–
A-3	Spleen	2007	134 (M)	+	+	+	+
A-4	Spleen	2008	4 (L)	+	–	+	+
A-5	Spleen	2009	97 (L)	+	–	+	+
A-6	Spleen	2010	32 (L)	+	–	–	+
A-7a	Kidney	2014	43 (L)	+	–	+	–
A-7b	Myocardium	2014	63 (L)	+	+	+	+
A-8	Liver	2014	439 (VH)	+	+	+	+
Clinical samples
A-9	CBS	2003	160 (M)	+	–	+	–
A-10	CBS	2007	78 (L)	+	+	+	+
A-11	CBS	2010	143 (M)	+	–	–	+
A-12	CBS	2011	112 (M)	+	–	–	+
A-13	CBS	2012	82 (L)	+	–	–	–

^aTissue used for cytological slide preparation
^bProtocol 1: conventional Babesia/Theileria PCR using BAB1 and BAB2 primers for amplification of 560 bp 18S rRNA; Protocol 2: conventional Babesia/Theileria PCR after DNA concentration and purification using BAB1 and BAB2 primers for amplification of 560 bp 18S rRNA; Protocol 3, nested Babesia/Theileria PCR for amplification of 1750 bp 18S rRNA and 560 bp 18S rRNA in the first and second round, respectively
Abbreviations: CBS, clinical blood smears made from the cephalic vein blood of clinical patients; L, low parasite burden; M, moderate parasite burden; H, high parasite burden; VH, very high parasite burden; cytb, mammalian cytochrome b

ISSN: 1756-3305