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Walz
Walz
KEY WORDS: Bovine viral diarrhea steers were exposed to the carcass of PI
virus, carrion transmission, Odocoileus fawn A (BVDV 2) or PI fawn B (BVDV 1).
virginianus, persistent infection, white-tailed Trials were designed with consideration of
deer the influence of contact networks on disease
epidemiology, and only one steer from each
ABSTRACT
group was separated into a pen with the
Infections with bovine viral diarrhea virus carcass. The number of contacts with the
(BVDV) clinically analogous to cattle are carcass was monitored. Following 8 hours,
described in white-tailed deer (Odocoileus the single steer was commingled with four
virginianus), but the epidemiologic role of other steers for 28 days. Animals were tested
persistently infected (PI) white-tailed deer is for BVDV infection. Controls included one
unknown. Persistently infected white-tailed steer inoculated intranasally with spleen-
deer shed BVDV, maintaining BVDV in homogenate from fawn A, and two steers
groups of deer. Survival of PI white-tailed inoculated intranasally or intravenously
deer is reduced, and clinically ill or dead PI with spleen-homogenate from fawn B.
deer may be a source of BVDV. This study Steers in both trials repeatedly contacted the
sought to determine if BVDV transmission carcasses, but BVDV transmission did not
could occur when cattle come in contact occur. The intranasally inoculated control
with carcasses of PI white-tailed deer. In for trial A and the intravenously inoculated
two trials, performed in Auburn, Alabama, control for trial B became viremic and
during November and December 2009, seroconverted. Although both PI carcasses
were potentially infectious and steers made born to experimentally infected does, had
repeated contact, transmission of BVDV did decreased survival and died before 10
not occur in this model. months of age.5 Early deaths of PI deer
reduce potential for BVDV transmission to
INTRODUCTION
susceptible animals by nose-to-nose contact,
Bovine viral diarrhea virus (BVDV), genus but infection of cattle following contact with
Pestivirus, family Flaviviridae, is a bovine dying or dead deer may be possible, and
pathogen, but the virus lacks host specificity, werethe subject of this investigation.
and infections occur in many Artiodactyls.1
Infection of free-ranging or captive wild MATERIALS AND METHODS
ruminants with BVDV may threaten natural Experimental Design and Animals
resources, including rare and endangered This research was performed under the
species. Contact of infected wildlife with approval of the Institutional Animal Care
susceptible cattle populations could pres- and Use Committee of Auburn University
ent a risk to ongoing BVDV control and (2009-1659). The study-design emulated
eradication programs.2 The risk of BVDV natural exposure of a susceptible steer to
transmission between wildlife and cattle is BVDV by presence of a deceased PI deer
currently unknown, but because wildlife and in the grazing area with consideration of
cattle commonly share habitat, this could influences of social hierarchies and contact
be a critical, yet poorly understood, aspect networks of cattle on disease transmission.11
of BVDV control programs in the United The study evaluated direct carcass-to-steer
States.3 transmission with subsequent transmission
BVDV infections clinically analogous to to herd-mates. This study consisted of two
bovine infections are reported in white-tailed separate trials, performed in November
deer (Odocoileus virginianus), and include and December 2009, each using one of two
reproductive losses and birth of persistent carcasses of PI fawns (Table 1). Fawns were
infections.4,5 Persistently infected (PI) white- born to dams experimentally exposed to
tailed deer can shed BVDV at similar levels either BVDV 2 strain PA131 (Fawn A) or
to PI cattle,6 and contact of pregnant does BVDV 1b strain AU526 (Fawn B) during
with PI fawns can result in BVDV transmis- gestation.12 A post-mortem examination
sion and birth of PI fawns.5 In the United was performed on the carcasses, which were
States, evidence of persistent infection in subsequently frozen at -20 °C for approxi-
free-ranging white-tailed deer was demon- mately 2 years until inclusion in this study.
strated, and reported prevalence rates for PI Two groups of five seronegative, BVDV-
white-tailed deer were 0.1 – 0.3%.7-10 negative Holstein steers were established for
Persistently infected white-tailed deer, carcass exposure experiments. Groups were
Fawn BVDV 0 hr 2 hr 4 hr 6 hr 8 hr
ID Swab Muscle Swab Swab Swab Swab Muscle
A Type 2 Virus + + + + + + +
PA131 isolation
Titrationa 6.2x103 2x103 BDTb 2x102 3.5x103 2x102 3.5x104
B Type 1 Virus - - - - - - -
AU526 isolation
Titrationa BDT BDT BDT BDT BDT BDT BDT
maintained in two-hectare pastures. Health tact with principal animals were inoculated
was assessed once daily. For each trial, the with spleen homogenates from fawns. One
carcass of a PI fawn was thawed at 5 °C for steer was intranasally inoculated with 2 ml
12 hours. One steer was separated from its of spleen homogenate in minimal essential
group into a pen of approximately 125 m2 medium (MEM) containing 1x104 CCID50/
with carcass exposure for 8 hours (Day 0). ml of BVDV from fawn A. Two steers were
Every 2 hours, the pen size was reduced by inoculated with 2 ml of inoculum containing
approximately half. Six hours after first ex- 0.25 ml of spleen homogenate from fawn B,
posure, feed was placed in direct vicinity of one intranasally, and the other intravenously.
the carcass. A contact with the carcass was While consistently positive by RT-PCR,
defined as the muzzle of the steer approach- virus isolation (VI) of tissues from carcass
ing the carcass at a distance of less than 20 B were negative at death and subsequent
cm. A separate event of contact was defined retesting, making quantification by virus
as being at least 15 minutes subsequent to titration impossible.
a prior event. The number of contacts of Samples Analyses
the steer with the carcass was monitored by Muscle samples were homogenized with a
time-lapse photography. Tekmar Stomacher (Model 80, Tekmar Co,
In order to limit behavioral alterations Cincinnati, OH, USA) and re-suspended
by presence of personnel, visual observa- with 3mL of MEM. White blood cells and
tions were not performed, except when sera were refrigerated for £ 72 hours before
samples were collected from the carcasses. VI procedures were performed. Sera were
To demonstrate infectious ability and evalu- removed and stored at -80º C for virus
ate survival of BVDV in tissues, two sites neutralization (VN) assays. To obtain the
in thoracic and abdominal cavities of each buffy coat, white blood cell samples were
carcass were swabbed with a Dacron polyes- processed as described previously,13 except
ter swab at 0, 2, 4, 6, and 8 hours. Additon- that samples were resuspended in 1 mL of
ally, muscle biopsy samples (1 cm3) were MEM.
collected from carcasses at 0 and 8 hours. Serum, buffy coat, carcass swab media,
Following 8 hours of exposure, the sepa- and muscle samples were assayed for BVDV
rated steer was returned to its group. Blood by passage though Madin Darby bovine kid-
samples for BVDV virology were collected ney (MDBK) cells for 3 days.13 The quantity
from steers on days 0, 4, 6 - 11, 14 - 18, 21, of BVDV in carcass swab media and muscle
22, and 28 of each trial. samples was determined by virus titration
Following the exposure trials, steer that performed in triplicate in 96-well plates con-
served as infection controls without con- taining MDBK cells. An immunoperoxidase