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Figure 1 Barranquilla working dog unit. evidence of tick-borne pathogens in three populations:
working dogs, shelter dogs, and client-owned/pet dogs.
For this study, point prevalence is defined as the pro-
portion of the Barranquilla canine population surveyed
that was positive for one or more tick-borne pathogens
at a specific point in time: July 2012. In this study, we
used species-specific peptide antigens to investigate the
seroprevalence of specific Ehrlichia spp. (E. canis, E.
chaffeensis, and E. ewingii) and specific Anaplasma spp.
(A. phagocytophilum and A. platys) in military work-
ing dogs, shelter animals, and client-owned pets in and
around the northern Colombian city of Barranquilla.
We also used nucleic acid–based molecular diagnostics
to elucidate the prevalence of rickettsemia present in the
Figure 2 Blood draw.
sample population.
Methods
Sample Population and Preparation
Blood samples (n = 218) were collected from client-
owned dogs presented to local veterinary clinics (n =
47), animals housed in regional community shelters
(n = 101), and from military working dogs (n = 70)
assigned to the area for duty. Blood was collected via
cephalic venipuncture directly into 2mL vacutainer
tubes with no anticoagulant for harvesting serum and
into 2mL vacutainer tubes containing ethylenediamine-
tetraacetic acid (EDTA) (Monoject ; Tyco, Mansfield,
™
MA, USA; http://www.tyco.com/). All study protocols
were approved through the University of Florida insti-
tutional animal care and use committee (protocol no.
201207460). Whole blood samples collected in EDTA
were stored at −20°C, while blood collected without
Figure 3 Map of Colombia. additives was immediately centrifuged; serum was har-
vested and was stored at −20°C. The samples were then
transported under cold conditions and stored at −20°C
in the laboratory of the investigators at the University
of Florida, College of Veterinary Medicine for analysis.
The samples were shipped in accordance with guidelines
Figure 4 Ticks under a paw. set forth in Permit no. 2012-05-164 from the Public
Health Service at the US Centers for Disease Control
and Prevention in Atlanta, GA, USA.
Enzyme-Linked Immunosorbent Assay
Serum samples were analyzed for the simultaneous de-
tection of antibodies against five different tick-borne
pathogens, Anaplasma phagocytophilum, A. platys,
Borrelia burgdorferi, Ehrlichia canis, and E. chaffeen-
sis using a research SNAP (IDEXX Laboratories, Inc.,
®
Westbrook, ME, USA; www.idexx.com) Multianalyte
Prototype Test. Briefly, three drops of patient serum
17
was mixed with four drops of conjugate and applied to
the flow matrix in an identical fashion as performed on
the commercially available ELISA assay SNAP 4Dx
®
®
Plus (IDEXX Laboratories, Inc., Westbrook, ME, USA;
82 Journal of Special Operations Medicine Volume 14, Edition 4/Winter 2014
