Two Anaplasma phagocytophilum Strains in Ixodes scapularis Ticks, Canada

Blacklegged ticks (n=12,606) were collected over four years (2007-2010) as part of a national passive tick surveillance program conducted in Canada by the National Microbiology Laboratory (NML) using the methods described previously (1). Adult I. scapularis were also collected by drag sampling: in 2008 from Itasca State Park, Camp Ripley and St. Croix State Park (n=56 per locality) in Minnesota (USA), in 2010 from within Pembina Valley Provincial Park (n=46), and along the Stanley Trail (n=44), in southern Manitoba, and in 2009 from the Wainfleet Bog Conservation area and St. Lawrence Islands National Park in southern Ontario (n=11). Total genomic (g) DNA was extracted from each tick at the NML or the University of Saskatchewan (U of S) using either the QIAamp DNA Mini Kit or DNeasy 96 Blood and Tissue Kit (QIAGEN, Ontario, Canada) as per the manufacturer’s instructions, but with the modifications described previously (1,2). The gDNA of A. phagocytophilum was also extracted (1) from an equine isolate (MN-93) propagated in the HL-60 promyelocytic cell line (ATCC CCL-240). This sample was used as a positive control in PCR assays conducted at the NML.