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Bovine Viral Diarrhea Virus Infections: Manifestations of Infection and Recent Advances in
Understanding Pathogenesis and Control
B. W. Brodersen
Vet Pathol 2014 51: 453 originally published online 29 January 2014
DOI: 10.1177/0300985813520250

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Review
Veterinary Pathology
2014, Vol. 51(2) 453-464
Bovine Viral Diarrhea Virus Infections: ª The Author(s) 2014
Reprints and permission:
sagepub.com/journalsPermissions.nav
Manifestations of Infection and Recent DOI: 10.1177/0300985813520250
vet.sagepub.com
Advances in Understanding Pathogenesis
and Control

B. W. Brodersen, DVM, MS, PhD1

Abstract
Bovine viral diarrhea virus (BVDV) continues to be of economic significance to the livestock industry in terms of acute disease and
fetal loss. Many of the lesions relating to BVDV infection have been well described previously. The virus is perpetuated in herds
through the presence of calves that are persistently infected. Relationships between various species and biotypes of BVDV and
host defenses are increasingly understood. Understanding of the host defense mechanisms of innate immunity and adaptive
immunity continues to improve, and the effects of the virus on these immune mechanisms are being used to explain how per-
sistent infection develops. The noncytopathic biotype of BVDV plays the major role in its effects on the host defenses by inhibiting
various aspects of the innate immune system and creation of immunotolerance in the fetus during early gestation. Recent advances
have allowed for development of affordable test strategies to identify and remove persistently infected animals. With these
improved tests and removal strategies, the livestock industry can begin more widespread effective control programs.

Keywords
cattle, domestic mammals, species, viral, infectious, disease, process, immunologic/inflammation, disease process, digestive tract,
tissue, lymphoreticular

Introduction and synthesis of costimulatory molecules.18,26,61,119 The

changes in cytokine production by immune or nonimmune cells
Bovine viral diarrhea virus (BVDV) is an economically impor-
affect both innate and adaptive immunity.108
tant pathogen that belongs to the genus Pestivirus of the family
The persistently infected (PI) animal serves as the reservoir
Flaviviridae.1 Bovine viral diarrhea virus causes bovine viral
of BVDV and a source of infection. Recent advances allow bet-
diarrhea (BVD), which was first reported as a transmissible ter detection of PI animals and thus improve the possibility of
disease in 1946.112 Since that time, BVD has been reported
enhanced control.
worldwide.46,50,128,136,147 Two genotypes, BVDV type 1 and
BVDV type 2, are further classified as cytopathogenic (cp)
or noncytopathogenic (ncp) based on in vitro cell culture Pathogenesis
characteristics.1,55,71,84,126
Two genotypes, BVDV type 1 and BVDV type 2, are identified
Modulation of the innate immune response in the fetus by
as distinct species within this genus, with further classification
ncp BVDV is of key importance in establishing persistent
as cp and ncp based on in vitro cell culture characteristics and
infection with BVDV. Understanding the interplay of BVDV
genetic differences.1,55,71,84,126 Genotype 1 is considered the
proteins and cellular regulation of the innate immune system
Pestivirus-type species and is reportedly the most prevalent
helps explain the development of persistent infection. Although
genotype.1,44,56
acute BVDV infections are often subclinical or produce only
mild clinical signs, they induce lymphopenia and a range of
effects on the immune response, which exacerbate secondary 1
Nebraska Veterinary Diagnostic Center, University of Nebraska–Lincoln,
bacterial or viral infections.19,79,89,117,119 BVDV infects a wide Lincoln, NE, USA
variety of cell types but has a predilection for cells of the
immune system such as monocytes/macrophages, dendritic Corresponding Author:
Bruce W. Brodersen, DVM, MS, PhD, Veterinary Diagnostic Center, School of
cells (DCs), and lymphocytes.19,61,90 The consequences of Veterinary Medicine and Biomedical Sciences, 1400 N 42nd St, University of
infection include the death of these cell populations as an Nebraska–Lincoln, Lincoln, Nebraska, USA.
extreme event or more subtle effects on cytokine expression Email: bbrodersen1@unl.edu

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454
There is evidence of immunologic cross-protection among
some strains of BVDV, including different genotypes.30,45,77,78
However, another study has shown incomplete immunologic
cross-protection between isolates and genotypes of BVDV.53
Type 2 genotypes are generally considered more virulent,
causing severe disease, including thrombocytopenia in some
cases.16,109 There is more likely a spectrum of virulence within
each Pestivirus species, and those spectra overlap. For exam-
ple, an outbreak in Spain was presumed to have been the result
of BVDV type 2 infection by veterinarians directly involved in
the outbreak,148 but later testing showed the cause to be a type
1b isolate, indicating that virulence of isolates can cross the
lines of what are perceived as low- and high-virulence BVDV
genotypes. One study demonstrated no difference in the
frequency of respiratory or enteric disease (or both) between
genotypes 1 and 2.56 Other reports describe high- and
low-virulence isolates of type 2 BVDV.29,90 A type 1b isolate,
New York 1, has been shown to be virulent and can cause leu-
kopenia, severe lymphoid depletion of thymus, mesenteric
lymph nodes, and Peyer patches as well erosions and ulcers
in the gastrointestinal tract.19 Thus, although type 2 BVDV is
generally thought to be more virulent, type 1 BVDV can cause
the same manifestations of disease.
Cattle are the natural host of the type species of pesti-
viruses,1 but infection by BVDV has been demonstrated in
numerous other species.5,43,58,63,76,145 Persistent infection with
BVDV is documented in alpacas, mouse deer, mountain goats,
white-tailed deer (WTD), sheep, and goats.5,8,43,106,131,145
Virus distribution among tissues in persistently infected sheep,
WTD, and mountain goats is similar to that seen in
cattle.72,106,115,127,131 The fact BVDV can infect a variety of
other species that are in natural contact with cattle poses a risk
for reinfection of pestivirus-susceptible cattle populations.83
Fetal infection may result in immunotolerance and lifelong
persistent infection.36,95,98,99 PI cattle with BVDV were
considered immunotolerant based on the fact they were serone-
gative for BVDV.98 It has been shown that persistent infection
does not result from cp BVDV infection; rather, infection with
ncp BVDV in early gestation will cause persistent infec-
tion.22,99 Persistently infected animals can develop severe
disease with widespread lesions of the alimentary mucosal
surfaces and lymphoid tissues, referred to as mucosal disease
(MD). Mucosal disease is the result of persistent infection with
a ncp strain of BVDV followed by subsequent postnatal infec-
tion with a cp strain of a closely related or homologous
virus.92,97,99 Homologous cp BVDV in an animal persistently
infected with ncp BVDV is thought to be the result of mutation
of that ncp BVDV. Induction of MD in cattle PI with BVDV
was not induced by administration of adrenocoticotropic
hormone, supporting the idea that MD was not a consequence
of glucocorticoid responses.85
Clinical signs of acute transient postnatal infection of animals
during the era when BVDV infection was emerging were
reported to be temperatures as high as 42 C, diarrhea, ulceration
of the muzzle and oral cavity, and leucopenia.112 Few or no clin-
ical signs were detected in other infected animals.112 This very
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Veterinary Pathology 51(2)
closely resembles clinical signs and lesions of animals with
MD that were seronegative for BVDV.111 Acute transient post-
natal infection remains one of the most important manifestations
of BVDV infection.29,52,89,94 Highly virulent strains of BVDV
can produce lesions similar to those seen in cases of MD, such
as severe and widespread ulceration of the oropharynx, larynx,
and esophagus and hemorrhagic enteritis.52,73,94,141 Observed
clinical signs consisted of inappetence, lethargy, and reduced
milk yield, with a spectrum of severity.
Abortion had been noted in several herds as BVDV was first
described and continues to be an economically important
aspect of BVDV infections in the present day.13,44,112 Fetal loss
due to in utero BVDV infection has been well recognized and is
often determined by the gestational age at the time of infec-
tion.28,100 Hormonal imbalances play a role in fetal loss, as
there are elevated prostaglandin levels in the dam during
BVDV infection that result in lysis of the corpus luteum (the
main source of progesterone in the cow).28
The lesions associated with acute BVDV infection were
described in the first reports. 112,144 Those lesions consisted
of erosions or ulcers in several tissues along the gastrointestinal
tract, ranging from oronasal (Fig. 1) erosions and ulcers, through
the pharynx, larynx (Fig. 2), esophagus (Fig. 3), forestomachs,
abomasum, and small and large intestines. Catarrhal enteritis
(Fig. 4) was described.121 Marked multifocal lymphoid deple-
tion, leaving karyorrhectic debris in germinal centers, can be
seen.144 Few lymphocytes are infected in Peyer patches com-
pared with the extent of lymphocyte destruction, suggesting an
indirect process is involved in depletion of lymphocytes.19,117
In acute infections, there can be lymphatic nodules in Peyer
patches that remain unaffected.80 Thymus is markedly depleted
of lymphocytes as the result of BVDV infection110 and can
be so severe as to leave just a collapsed stroma with few
hemorrhages and scattered lymphocytes.19 Some lymphoid
lesions were mild despite the presence of large amounts
of viral antigen in calves infected with low-virulence
virus.91 Viral antigen is widely distributed throughout
lymphoid tissues such as the palatine tonsil, thymus, and
Peyer patches.19,80,88,111 BVDV antigen is demonstrated in
interdigitating cells in lymphoid tissues during both acute
and persistent infection.12,19
Comparison of clinically normal PI cattle with calves
acutely (transiently) infected by the same virus was reported.89
Viral antigen was reported in all tissues of the PI calves and
was not associated with lesions. In the acutely infected animals,
viral antigen was found mainly between days 5 and 9 postino-
culation. Viral antigen was noted mainly in lymphoid tissues
but not bone marrow. Other tissues were infected in only a few
of the animals on various days during the 5- to 9-day period.
Clinical signs were mild in these calves, and there was no
thrombocytopenia. Lymphoid depletion was mild at day 3 post-
infection, with increasing severity until day 9, and by day 13
there was evidence of repopulation of lymphoid tissues. This
was consistent with what others have reported.19,77,80 Severe
fibrinoid necrosis of blood vessel walls was seen in lymph
nodes and heart of 1 calf on day 13 postinoculation.89
Brodersen 455

Figures 1–6. Lesions in cattle due to bovine viral diarrhea virus infection. Figure 1. Planum nasale, bovine. Severe diffuse ulcerative epidermitis.
Figure 2. Larynx, bovine. Severe diffuse ulceration of mucosa of arytenoid cartilages and pharyngoesophageal junction. Figure 3. Esophagus,
bovine. Multiple mucosal ulcers. Figure 4. Jejunum, bovine. Necrosis of jejunal Peyer patch and catarrhal enteritis. Figure 5. Heart, bovine.
Myocardial degeneration and necrosis. Hematoxylin and eosin (HE). Figure 6. Heart, bovine. Serial section from Fig. 5, intralesional positive
BVDV staining. Immunohistochemistry, hematoxylin counterstain.

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456
Myocardial necrosis has been seen in 8-month-old calves with
BVDV antigen associated with the lesion (personal observa-
tion) (Figs. 5, 6), although it was not known if these were PI
or acutely infected animals.
Thus, the patterns of distribution of viral antigen differ in
acute transient postnatal infection, persistent infection, and
MD.88 In acute transient postnatal infection, there is highly
variable antigen distribution with marked depletion of lymphoid
follicles in several tissues.80,88 In PI, there is widespread distri-
bution of viral antigen, including skin and salivary glands, but
only mild or locally restricted depletion of lymphoid tissues.111
In MD, widespread distribution of viral antigen including the
salivary glands, severe depletion of all lymphoid tissues, and
severe lesions in the gastrointestinal tract are observed.88
Special Pathology
In addition to the gastrointestinal tract and lymphoid tissues,
BVDV antigens have been demonstrated in a variety of
tissues, including the cerebellum of mid-gestation fetuses,
blood vessel walls, integument, and placenta from aborted
fetuses.48,110,111,120,123 Hypoplastic lesions of the cerebellum,
affecting all folia, are described in a calf whose dam was
infected as early as 79 days of gestation, and lesions of cavita-
tion, folial degeneration, and granule cell depletion were present
in calves from dams infected later in gestation up to 150 days.21
Neurotropic strains of ncp BVDV, which caused multifocal
meningoencephalitis with neuronal degeneration and necrosis,
have been reported.14,47,76,104 In experimental infection of lambs
and calves, low- and high-virulence strains differ in the resulting
clinical and pathological findings, as well as in the amount of
viral antigen present.76,80,91 Following experimental infection
of pregnant cows at 75 days of gestation, viral antigen was wide-
spread in the brain of the fetuses, particularly in nuclei in the tha-
lamus. These findings were similar to natural infection.103,104
Lesions of leukomalacia and infiltration of the meninges and
neuropil by macrophages and neutrophils were seen.11,104 The
fetal brain is infected via infiltrating microglial precursor cells
(ameboid glial cells), and vasculopathy occurs around the time
of viral invasion. This is followed by microvascular loss (pre-
sumably by necrosis), which either leads to or is concurrent with
microcavity formation or cysts that persist and may enlarge over
subsequent weeks.10 It appears neuronal infection is abortive, as
viral antigen appears restricted to the Golgi region of most
infected neurons.10
Ocular lesions in fetuses 17 to 22 days after experimental
mid-gestational infection of cows consisted of mild to moder-
ate retinitis, mononuclear cuffing of inner retinal vessels,
proliferation of pigment epithelium, and choroiditis.20 Retinal
atrophy was observed in newborn calves that were infected in
mid-gestation.20 Congenital cataracts have been reported in
calves with BVDV infection, but it is unclear as to the circum-
stances of the infections relative to early or late gestation or
characteristics of the infecting BVDV.151
Bovine viral diarrhea virus has been associated with growth
retardation lattices of long bones, described as sclerotic lines of
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Veterinary Pathology 51(2)
metaphyseal bone parallel to the physis.142 Experimental
infection of pregnant cows at 75 days of gestation resulted in
altered metaphyseal bone density, first detected at 192 days
of gestation.149 It was hypothesized that in the primary spon-
giosa, a lesser amount of residual intercolumnar cartilage was
resorbed before osteoid formation commenced.149 However,
there was no evidence of reduced numbers of osteoclasts
compared with postnatally infected calves, in which osteoclasts
were not seen in primary or secondary spongiosa.135
Chronic postnatal infections exist in animals in ‘‘immuno-
privileged’’ sites such as the testicle and ovary. Aside from the
impact of BVDV on fetuses in terms of congenital defects and
abortions, acute BVDV infection can have a direct impact on
reproductive performance.59 As a result of acute infection,
there can be prolonged testicular infection and shedding of
virus in semen for as long as 5 months after infection.59 BVDV
has been demonstrated in cells lining seminiferous tubules.59
In the ovaries of PI heifers, BVDV has been demonstrated in
cumulus cells, oocytes in all stages of maturation, and ovarian
stroma.51 Oophoritis, characterized by the presence of virus-
infected macrophages, was seen following acute infection of
heifers.66 Follicular growth in ovaries from acutely infected
heifers was impaired during the 2 subsequent estrous cycles
after acute infection.65
Persistent Infection
Persistent infection is induced by infection of the fetus with ncp
BVDV early in gestation (40–125 days) before development of
the humoral immune system.36,98,99 To persist, both the innate
and adaptive immune responses are averted by the virus.
Type I interferons (IFNs) are important mediators of the
innate immune responses to viruses, and BVDV interference
with type I IFN signaling has been implicated as a contributing
factor in the establishment of PI.32,133,137 Macrophages
infected with ncp BVDV (of relevance to persistent infection)
fail to induce type I IFN, whereas macrophages infected with
cp BVDV (of relevance to mucosal disease) do release type I
IFN.2,3,31,32,118,133 Similarly, infection of 60-day bovine fetuses
with ncp BVDV did not result in detectable IFN-a/b in amnio-
tic fluid, whereas IFN-a/b was detected in amniotic fluid
following infection with cp BVDV.17,32
Viral double-stranded and single-stranded RNA
fragments are able to stimulate type I IFN production.96
Double-stranded RNA is able to stimulate production of type
I IFN in a RIG-I–independent manner.96,146 RIG-I detects
double-stranded RNA during replication of positive sense
single-stranded RNA, which leads to activation of interferon
regulatory factor 3 (IRF-3).150 In addition, Toll-like receptor
(TLR) 3 binds to double-stranded RNA and activates IRF-
3.150 Viral RNA is recognized by pattern recognition receptors
such as TLRs.87 Cells infected with cp BVDV contain more
viral double-stranded RNA (a replication intermediate of many
RNA viruses) than cells infected with ncp BVDV.96 The
greater amount of dsRNA produced by cp BVDV has been
postulated to contribute to stimulation of IFN production via
Brodersen
RIG-I– and TLR-3–mediated activation of IRF-3.96 Conver-
sely, the BVDV glycoprotein Erns is able to inhibit IFN expres-
sion, and the RNAse activity of Erns is required for this
inhibition.96 Erns lacks a typical transmembrane anchor and
is not tightly bound to cellular membranes,49 so this RNAse
activity of Erns is able to degrade extracellular single- and
double-stranded RNA. Since these are triggers of type I IFN
production, this is one way that Erns prevents type I IFN induc-
tion even in uninfected cells.96
The BVDV glycoprotein Npro has been shown to be essential in
blocking induction of type I IFN. Involved in the IFN-regulatory
cascade are IRF-3 and IRF-7. IRF-3 is depleted by the viral pro-
tein Npro through proteasomal degradation.6 The combined effect
of Npro and Erns is necessary to adequately block IFN production
and establish PI. Infection of fetuses with Npro- or Erns-deleted
mutants of ncp BVDV alone was not sufficient to block establish-
ment of fetal infection. Mutants of ncp BVDV containing both
deletions blocked fetal infection in vivo.101
Additional work has conversely demonstrated there is
activation of the innate response in dams of calves in the first
trimester as a result of ncp BVDV infection,139 but this is only
partially effective in curtailing viral replication, and an adap-
tive response is required to eliminate infection. Upregulation
of interferon-stimulated gene 15kD (ISG15) has been observed
in the blood of pregnant cattle during the early stage of ncp
BVDV infection both in early and middle gestation.137,138 By
day 45 postinoculation, ISG15 had returned to baseline in both
the PI and transiently infected (TI) cows in PI mothers, using
quantitative reverse transcription polymerase chain reaction
(qRT-PCR). Further work has shown that ncp BVDV has
effects on several families of chemokines.27 There is upregula-
tion of type I IFN in maternal blood of calves infected with ncp
BVDV at day 175 of gestation.138 Leukocytes from these dams
with a PI fetus showed downregulation of T-cell receptor
signaling and of CXCR4 (the receptor for the lymphocyte che-
moattractant, stromal cell–derived factor 1).15,123,138 It is these
features of the BVDV proteins Erns and Npro that are thought to
enable the virus to avoid the innate immune system in the early
gestation fetus.
Acute Infection
Little has been published regarding innate immunity to BVDV
in the postnatal animal. Generally, the innate response to
BVDV in the postnatal animal is not considered unique among
viral infections. However, apparent differences exist between
high-virulence (HV) and low-virulence (LV) isolates as well
as between cp BVDV and ncp BVDV. Infection of 7-month-
old calves with a high-virulence ncp BVDV isolate has been
shown to upregulate interferon a and b messenger RNA
(mRNA) levels in tracheobronchial lymph nodes in the HV
group but not in the LV group.114 Type I IFN genes in spleens
of both HV and LV compared with control calves were
upregulated.114
Important differences occur between ncp BVDV and cp
BVDV in the induction of apoptosis. Cytopathic BVDV, but
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457
not noncytopathic (ncp) BVDV, has been shown to induce
apoptosis in vitro in embryonic bovine turbinate cells.68,134,154
Infection of several types of cultured cells with the cp, but not
with the ncp, biotype of BVDV triggers the induction of
apoptosis.2 Expression of the NS3 protease of cp BVDV is
unable to inhibit TLR-3– and RIG-I–dependent activation of
IFN-b, thus allowing apoptosis to occur in cp BVDV infections.57
Whereas the viral protease NS3 is continually expressed by
cp BVDV-infected cells and is thought to be important in indu-
cing apoptosis, this protein is expressed only early in cellular
infection or not at all in ncp BVDV infections. This difference
may partially explain the pathologic findings of cell death
observed in mucosal disease resulting from cp BVDV infec-
tion, whereas such lesions are not observed in calves persis-
tently infected with ncp BVDV.
There are conflicting reports regarding apoptosis due to ncp
BVDV infection. The intrinsic pathway is induced, at least in
vitro, by ncp BVDV by overexpression of apoptotic protease-
activating factor 1 and increasing caspase 9 activity.67 Further
evidence supporting the intrinsic pathway as a mechanism for
apoptosis during acute BVDV infection is a reduction in the
number of tumor necrosis factor–a (TNF-a)–expressing cells
in the thymus of calves experimentally inoculated with a type
1 ncp BVDV.122 Noncytopathic BVDV can prime bone mar-
row–derived macrophages for production of reactive nitrogen
in response to stimulation,3 and reactive nitrogen (nitric oxide)
can induce the extrinsic pathway of apoptosis.24
BVDV can induce apoptosis of lymphocytes through the
RNase activity of Erns.25,70,132 In cells infected with ncp
BVDV, the NS2/3 protein is cleaved in the early stages of
cellular infection, to release the proapoptotic viral protein
NS3. That cleavage is then reduced, resulting in the presence
of predominantly uncleaved NS2/3 in later stages of infec-
tion.84 The viral protein NS3 induces apoptosis via activation
of caspase 8 and the cytochrome c release-dependent caspase
9, thus inducing both extrinsic and intrinsic pathways.140
Induction of moderate amounts of caspase 8 is associated with
apoptosis of T-lymphocyte–rich interfollicular areas and
marked apoptosis of B lymphocytes in lymphoid follicles dur-
ing infection of ileal Peyer patches.116 This finding suggests a
role of the extrinsic pathway of apoptosis in gut-associated
lymphoid tissue (GALT).
Interestingly, it was demonstrated there is inactivation of
caspase 9 during acute infection with ncp BVDV in GALT,
suggesting the intrinsic pathway is not involved in apoptosis
during ncp BVDV infection.116 Further evidence that the
intrinsic pathway is regulated during ncp BVDV infection is
that reports of the apoptotic pathway regulator, Bcl-2, blocks
the actions of the Bax gene in a balance between Bcl-2 and
Bax.23,38,102 The antiapoptotic Bcl-2 protein is upregulated
by ncp BVDV, suppressing initiation of intrinsic apoptosis
and cell death.9 Overexpression of Bcl-2 in T-cell interfollicu-
lar areas may have an inhibitory effect on apoptosis, protecting
T lymphocytes in those interfollicular areas.116 Reduced levels
of Bcl-2 in follicular areas during acute infection could be asso-
ciated with apoptosis of B lymphocytes.116 Oxidative stress
458
occurs early in the interaction between cp BVDV and its host
cell and may be a crucial event in the sequence leading to
apoptotic cell death.134 Hence, apoptosis is not required for the
multiplication of the cp biotype of BVDV.134
Adaptive Immunity, Vaccination, and
Immunosuppression
Modified live and killed vaccines have been available for more
than 50 years, yet the incidence of BVDV-induced disease
remains significant, suggesting a need for improved vaccines.
This may be due, in part, to the lack of proofreading function
during replication of the single-stranded genome of BVDV and
the resultant antigenic variability.124 Neutralizing antibodies
prevent disease following challenge with homologous virus,
as well as prevent or ameliorate disease with challenge with
heterologous virus.23,30,53,54 There are 4 structural proteins of
BVDV, but the E2 glycoprotein is the major target of neutraliz-
ing antibodies.40,41 Newly designed vaccines have addressed
the issues related to virulence of E2-expressing vaccine viruses
by incorporating the E2 gene into nonvirulent vectors.4,93 The
E2 region of the genome has been used as one of the regions for
classification of BVDV isolates due to its variability in the
genome sequence.7,125,148 Due to the antigenic variability,
there is potential for a lack of cross-protection against wild-
type BVDV viruses, even though cross-neutralization studies
have shown reactivity to differing genotypes.93 Since disease
due to BVDV remains widespread, additional measures beyond
vaccination are needed.
Resolution of postnatal infection is reliant on function of
CD4þ T cells,75 and one of the mechanisms for persistence
is tolerance of these cells due to in utero infection.34 It has been
shown that as little as 1 amino acid difference in quasispecies is
sufficient to stimulate an antibody response to homologous
virus, demonstrating the specificity of the tolerance of the
adaptive immune response.34
Vaccines are not without their adverse effects on the host. In
the event that there is PI in animals with BVDV in a group that
is administered a closely related cp BVDV in the modified live
vaccine virus, there is potential for the unfortunate occurrence
of MD as a result of these combined infections.125 Modified
live vaccines for BVDV can retain some virulence and can
result in lymphoid depletion in Peyer patches as well as altera-
tion of neutrophil and lymphocyte functions after vaccina-
tion.78,129 Vaccines can also be contaminated by virulent
BVDV, resulting in PI in calves with BVDV after vaccination
of pregnant animals.113 Vaccine virus can also be shed after
extra-label administration to pregnant animals and, in the
proper circumstances, can result in PI calves.113
Immunosuppression is an important feature of BVDV infec-
tion. Both monocytes and DCs are susceptible to infection by
cp and ncp BVDV.61 Monocytes but not DCs were killed by
cp BVDV, while ncp BVDV infection reduced responses of
CD4þ T cells to antigen.61 Macrophages infected with ncp
BVDV have downregulation of CC- and CXC-chemokine
expression.27 During acute infections, antigen-presenting cells
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Veterinary Pathology 51(2)
are compromised, contributing to immune suppression. Marked
depletion of CD4þ and CD8þ cells has been reported in the
thymus.18 In vivo infection with ncp BVDV reduced the num-
ber of major histocompatability complex (MHC) class II cells
in Peyer patches and lymph nodes.18 Infection of peripheral
blood mononuclear cells with cp BVDV results in altered
expression of proteins involved in adhesion, apoptosis, antigen
uptake, processing, and presentation, thus compromising the
response of antigen-presenting cells.86 However, monocytes
from calves with PI are nonetheless able to generate MHC class
I–restricted and MHC class II–restricted responses to BVDV.62
Contemporary Diagnostic Methods
Several techniques for the diagnosis of BVDV infections have
been developed recently that have allowed affordable wide-
spread testing of animals for BVDV infection. Those tech-
niques have included both immunoenzymatic assays as well
as PCR assays. Early immunohistochemical tests were based
on cryosections of fresh tissues.64,143,152 However, later tech-
niques were developed, using formalin-fixed, paraffin-
embedded (FFPE) tissues.69 This was based on the monoclonal
antibody directed against the Erns glycoprotein of BVDV,41
which was shown to recognize all isolates of BVDV tested.35
This same MAb was used in the development of an antigen
capture enzyme-linked immunosorbent assay (ELISA).130
The PI animal serves as a reservoir for BVDV and is a
source of infection for herd mates. Control strategies have
focused on identification and removal of PI animals from
herds. Previously, PI animals were identified by repeated
demonstration of virus in buffy coat cells or nasal or lacrimal
exudate from the same animal.98 These animals also remain
seronegative for extended periods. Contrasted to this, acutely
infected animals remain viremic for a relatively short period
(generally less than 3 weeks) and seroconvert, producing
neutralizing antibodies as early as 2 to 3 weeks after infec-
tion.77,98,117 In acutely infected animals, virus is cleared from
most tissues by day 9 postinfection, and these animals gener-
ally do not serve as a source of infection to herd mates. 89 Stra-
tegies to control BVDV infection in herds include vaccination
and removal of PI animals. The prevalence of PI animals is
reported to be between 1% and 2%,74,153 so large numbers of
animals need to be tested to identify those few that have PI with
BVDV. As a result, it is important to be able to affordably iden-
tify the PI animal. Virus isolation was too slow and expensive
for large-scale testing. The immunoperoxidase microplate virus
isolation test was developed for serum, which greatly enhanced
screening for PI animals.39
In early work, skin was identified as a location of viral anti-
gen, and subsequently immunohistochemistry on cryosections
of skin biopsies was shown to reliably identify PI animals with
BVDV.111,143 Use of skin along with development of immuno-
histochemistry (IHC) on FFPE tissues enabled high-throughput
testing for identification of PI animals. That is when the idea of
using FFPE ear notch samples was developed (Brodersen, Pro-
ceedings 31st AABP Annual Meeting, 1998. Spokane, WA).
Brodersen
Additional testing methodologies have used ear notches as the
substrate—namely, antigen capture ELISA and
PCR.37,82,105,107 In large-scale outbreaks of BVDV with many
PI animals, BVDV antigen is expressed in the skin of transi-
ently infected animals for as long as 120 days.37 In those situa-
tions, it is advisable to use different methodologies over an
extended period to accurately identify which animals are PI and
which are transiently infected. It is particularly important to
perform additional testing on animals suspected of being PI
when blood samples and nasal swabs are used during initial
testing.54 The current testing methodologies allow for imple-
mentation of BVDV control strategies.42
Summary
BVDV is a pestivirus that can cause severe clinical disease.
Major economic losses continue to result from reproductive
losses and exacerbation of concurrent bacterial or viral infec-
tions. Of less clinical importance is severe clinical disease due
to BVDV alone, either as a result of acute infection by highly
virulent isolates or by development of mucosal disease in
persistently infected calves. Disease due to BVDV infection
can arise at any stage of life and is often dependent on the viru-
lence of the infecting isolate.
Survival of BVDV in the population depends on the charac-
teristic of persistently infecting its host. Persistent infection
arises by the unique ability of the BVDV to survive by inducing
immune tolerance in the bovine fetus through evasion of both
innate and acquired immunity in utero. Adaptive immunity is
avoided by immune tolerance through infection of the fetus
prior to development and maturation of the adaptive immune
system. The PI calf with BVDV then serves as a source of
infection of herd mates and thereby leads to continuation of the
viral species. Because of this unique and complex pathogen-host
interaction, a voluminous body of research has been produced
and summarized here as well as in other sources.33,55,60,81 Con-
trol of BVDV infections in herds is dependent on identification
of PI calves with BVDV and removal of those PI animals.
Acknowledgement
I wish to thank Dr. Clayton L. Kelling for his advise in composition of
this manuscript.
Declaration of Conflicting Interests
The author(s) declared no potential conflicts of interest with respect to
the research, authorship, and/or publication of this article.
Funding
The author(s) received no financial support for the research, author-
ship, and/or publication of this article.
References
1. Virus Taxonomy: Classification and Nomenclature of Viruses:
Ninth Report of the International Committee on Taxonomy of
Viruses. San Diego, CA: Elsevier/Academic Press; 2012.
Downloaded from vet.sagepub.com at UNIV ARIZONA LIBRARY on June 15, 2014
459
2. Adler B, Adler H, Pfister H, et al. Macrophages infected with
cytopathic bovine viral diarrhea virus release a factor(s) capable
of priming uninfected macrophages for activation-induced apop-
tosis. J Virol. 1997;71:3255–3258.
3. Adler H, Frech B, Meier P, et al. Noncytopathic strains of bovine
viral diarrhea virus prime bovine bone marrow–derived macro-
phages for enhanced generation of nitric oxide. Biochem Biophys
Res Commun. 1994;202:1562–1568.
4. Aguirreburualde MS, Gomez MC, Ostachuk A, et al. Efficacy of a
BVDV subunit vaccine produced in alfalfa transgenic plants. Vet
Immunol Immunopathol. 2013;151:315–324.
5. Bachofen C, Vogt HR, Stalder H, et al. Persistent infections after
natural transmission of bovine viral diarrhoea virus from cattle to
goats and among goats. Vet Res. 2013;44:32.
6. Bauhofer O, Summerfield A, Sakoda Y, et al. Classical swine
fever virus Npro interacts with interferon regulatory factor 3 and
induces its proteasomal degradation. J Virol. 2007;81:3087–3096.
7. Becher P, Orlich M, Kosmidou A, et al. Genetic diversity of
pestiviruses: identification of novel groups and implications for
classification. Virology. 1999;262:64–71.
8. Bedenice D, Dubovi E, Kelling CL, et al. Long-term clinicopatho-
logical characteristics of alpacas naturally infected with bovine
viral diarrhea virus type Ib. J Vet Intern Med. 2011;25:605–612.
9. Bendfeldt S, Grummer B, Greiser-Wilke I. No caspase activation
but overexpression of Bcl-2 in bovine cells infected with noncy-
topathic bovine virus diarrhoea virus. Vet Microbiol. 2003;96:
313–326.
10. Bielefeldt-Ohmann H, Smirnova NP, Tolnay AE, et al. Neuro-
invasion by a ‘Trojan Horse’ strategy and vasculopathy during
intrauterine flavivirus infection. Int J Exp Pathol. 2012;93:24–33.
11. Bielefeldt-Ohmann H, Tolnay AE, Reisenhauer CE, et al. Trans-
placental infection with non-cytopathic bovine viral diarrhoea
virus types 1b and 2: viral spread and molecular neuropathology.
J Comp Pathol. 2008;138:72–85.
12. Bielefeldt Ohmann H. BVD virus antigens in tissues of persis-
tently viraemic, clinically normal cattle: implications for the
pathogenesis of clinically fatal disease. Acta Vet Scand. 1988;
29:77–84.
13. Blanchard PC, Ridpath JF, Walker JB, et al. An outbreak of
late-term abortions, premature births, and congenital deformities
associated with a bovine viral diarrhea virus 1 subtype b that
induces thrombocytopenia. J Vet Diagn Invest. 2010;22:128–131.
14. Blas-Machado U, Saliki JT, Duffy JC, et al. Bovine viral diarrhea
virus type 2–induced meningoencephalitis in a heifer. Vet Pathol.
2004;41:190–194.
15. Bleul CC, Fuhlbrigge RC, Casasnovas JM, et al. A highly
efficacious lymphocyte chemoattractant, stromal cell–derived
factor 1 (SDF-1). J Exp Med. 1996;184:1101–1109.
16. Bolin SR, Ridpath JF. Differences in virulence between two
noncytopathic bovine viral diarrhea viruses in calves. Am J Vet
Res. 1992;53:2157–2163.
17. Brackenbury LS, Carr BV, Charleston B. Aspects of the innate
and adaptive immune responses to acute infections with BVDV.
Vet Microbiol. 2003;96:337–344.
18. Brodersen BW, Kelling CL. Alteration of leukocyte populations
in calves concurrently infected with bovine respiratory syncytial
460
virus and bovine viral diarrhea virus. Viral Immunol. 1999;12:
323–334.
19. Brodersen BW, Kelling CL. Effect of concurrent experimentally
induced bovine respiratory syncytial virus and bovine viral diar-
rhea virus infection on respiratory tract and enteric diseases in
calves. Am J Vet Res. 1998;59:1423–1430.
20. Brown TT, Bistner SI, de Lahunta A, Scott FW, et al. Pathoge-
netic studies of infection of the bovine fetus with bovine
viral diarrhea virus, II: ocular lesions. Vet Pathol. 1975;12:
394–404.
21. Brown TT, DeLahunta A, Bistner SI, et al. Pathogenetic studies of
infection of the bovine fetus with bovine viral diarrhea virus, I:
cerebellar atrophy. Vet Pathol. 1974;11:486–505.
22. Brownlie J, Clarke MC, Howard CJ. Experimental infection of
cattle in early pregnancy with a cytopathic strain of bovine virus
diarrhoea virus. Res Vet Sci. 1989;46:307–311.
23. Brun A, Rivas C, Esteban M, et al. African swine fever virus gene
A179 L, a viral homologue of bcl-2, protects cells from pro-
grammed cell death. Virology. 1996;225:227–230.
24. Brune B.Nitric oxide: NO apoptosis or turning it ON? Cell Death
Differ. 2003;10:864–869.
25. Bruschke CJ, Hulst MM, Moormann RJ, et al. Glycoprotein Erns
of pestiviruses induces apoptosis in lymphocytes of several
species. J Virol. 1997;71:6692–6696.
26. Buckwold VE, Wei J, Wenzel-Mathers M, et al. Synergistic in
vitro interactions between alpha interferon and ribavirin against
bovine viral diarrhea virus and yellow fever virus as surrogate
models of hepatitis C virus replication. Antimicrob Agents
Chemother. 2003;47:2293–2298.
27. Burr S, Thomas C, Brownlie J, et al. Potential evidence for
biotype-specific chemokine profile following BVDV infection
of bovine macrophages. Vet Immunol Immunopathol. 2012;150:
123–127.
28. Carlsson U, Fredriksson G, Alenius S, et al. Bovine virus
diarrhoea virus, a cause of early pregnancy failure in the cow.
Zentralbl Veterinarmed A. 1989;36:15–23.
29. Carman S, van Dreumel T, Ridpath J, et al. Severe acute bovine
viral diarrhea in Ontario, 1993–1995. J Vet Diagn Invest. 1998;
10:27–35.
30. Castrucci G, Avellini G, Cilli V, et al. A study of immunologic
relationships among serologically heterologous strains of bovine
viral diarrhea virus by cross immunity tests. Cornell Vet. 1975;
65:65–72.
31. Charleston B, Brackenbury LS, Carr BV, et al. Alpha/beta and
gamma interferons are induced by infection with noncytopathic
bovine viral diarrhea virus in vivo. J Virol. 2002;76:923–927.
32. Charleston B, Fray MD, Baigent S, et al. Establishment of persis-
tent infection with non-cytopathic bovine viral diarrhoea virus in
cattle is associated with a failure to induce type I interferon. J Gen
Virol. 2001;82:1893–1897.
33. Chase CC. The impact of BVDV infection on adaptive immunity.
Biologicals. 2013;41:52–60.
34. Collen T, Douglas AJ, Paton DJ, et al. Single amino acid differ-
ences are sufficient for CD4(þ) T–cell recognition of a heterolo-
gous virus by cattle persistently infected with bovine viral
diarrhea virus. Virology. 2000;276:70–82.
Downloaded from vet.sagepub.com at UNIV ARIZONA LIBRARY on June 15, 2014
Veterinary Pathology 51(2)
35. Corapi WV, Donis RO, Dubovi EJ. Monoclonal antibody analy-
ses of cytopathic and noncytopathic viruses from fatal bovine
viral diarrhea virus infections. J Virol. 1988;62:2823–2827.
36. Coria MF, McClurkin AW. Specific immune tolerance in an
apparently healthy bull persistently infected with bovine viral
diarrhea virus. J Am Vet Med Assoc. 1978;172:449–451.
37. Cornish TE, van Olphen AL, Cavender JL, et al. Comparison of
ear notch immunohistochemistry, ear notch antigen-capture
ELISA, and buffy coat virus isolation for detection of calves
persistently infected with bovine viral diarrhea virus. J Vet Diagn
Invest. 2005;17:110–117.
38. Cory S, Huang DC, Adams JM. The Bcl-2 family: roles in cell
survival and oncogenesis. Oncogene. 2003;22:8590–8607.
39. Deregt D, Prins S. A monoclonal antibody-based immunoperoxi-
dase monolayer (micro-isolation) assay for detection of type 1 and
type 2 bovine viral diarrhea viruses. Can J Vet Res. 1998;62:
152–155.
40. Donis RO. Molecular biology of bovine viral diarrhea virus and
its interactions with the host. Vet Clin North Am Food Anim Pract.
1995;11:393–423.
41. Donis RO, Corapi W, Dubovi EJ. Neutralizing monoclonal
antibodies to bovine viral diarrhoea virus bind to the 56 K to 58
K glycoprotein. J Gen Virol. 1988;69(pt 1):77–86.
42. Dubovi EJ. Laboratory diagnosis of bovine viral diarrhea virus.
Biologicals. 2013;41:8–13.
43. Duncan C, Ridpath J, Palmer MV, et al. Histopathologic and
immunohistochemical findings in two white-tailed deer fawns
persistently infected with bovine viral diarrhea virus. J Vet Diagn
Invest. 2008;20:289–296.
44. Evermann JF, Ridpath JF. Clinical and epidemiologic observa-
tions of bovine viral diarrhea virus in the northwestern United
States. Vet Microbiol. 2002;89:129–139.
45. Fairbanks K, Schnackel J, Chase CC. Evaluation of a modified
live virus type-1a bovine viral diarrhea virus vaccine (Singer
strain) against a type-2 (strain 890) challenge. Vet Ther. 2003;4:
24–34.
46. Falcone E, Cordioli P, Sala G, et al. Genotyping of bovine viral
diarrhoea viruses isolated from cattle in northern Italy. Vet Res
Commun. 2001;25:161–167.
47. Fernandez A, Hewicker M, Trautwein G, et al. Viral antigen dis-
tribution in the central nervous system of cattle persistently
infected with bovine viral diarrhea virus. Vet Pathol. 1989;26:
26–32.
48. Fernelius AL, Lambert G. Detection of bovine viral diarrhea virus
and antigen in tissues of experimentally infected calves by cell
inoculation and fluorescent antibody techniques. Am J Vet Res.
1969;30:1551–1559.
49. Fetzer C, Tews BA, Meyers G. The carboxy-terminal sequence of
the pestivirus glycoprotein E(rns) represents an unusual type of
membrane anchor. J Virol. 2005;79:11901–11913.
50. Flores EF, Ridpath JF, Weiblen R, et al. Phylogenetic analysis of
Brazilian bovine viral diarrhea virus type 2 (BVDV-2) isolates:
evidence for a subgenotype within BVDV-2. Virus Res. 2002;
87:51–60.
51. Fray MD, Prentice H, Clarke MC, et al. Immunohistochemical
evidence for the localization of bovine viral diarrhea virus, a
Brodersen
single-stranded RNA virus, in ovarian oocytes in the cow. Vet
Pathol. 1998;35:253–259.
52. Friedgut O, Rotenberg D, Brenner J, et al. Description of the first
acute bovine diarrhea virus-2 outbreak in Israel. Vet J. 2011;189:
108–110.
53. Fulton RW, Briggs RE, Ridpath JF, et al. Transmission of bovine
viral diarrhea virus 1b to susceptible and vaccinated calves by
exposure to persistently infected calves. Can J Vet Res. 2005;
69:161–169.
54. Fulton RW, Hessman BE, Ridpath JF, et al. Multiple diagnostic
tests to identify cattle with bovine viral diarrhea virus and dura-
tion of positive test results in persistently infected cattle. Can J
Vet Res. 2009;73:117–124.
55. Fulton RW, Ridpath JF, Confer AW, et al. Bovine viral diarrhoea
virus antigenic diversity: impact on disease and vaccination pro-
grammes. Biologicals. 2003;31:89–95.
56. Fulton RW, Ridpath JF, Ore S, et al. Bovine viral diarrhoea virus
(BVDV) subgenotypes in diagnostic laboratory accessions: distri-
bution of BVDV1a, 1b, and 2a subgenotypes. Vet Microbiol.
2005;111:35–40.
57. Gamlen T, Richards KH, Mankouri J, et al. Expression of the NS3
protease of cytopathogenic bovine viral diarrhea virus results in
the induction of apoptosis but does not block activation of the beta
interferon promoter. J Gen Virol. 2010;91:133–144.
58. Gao Y, Wang S, Du R, et al. Isolation and identification of a
bovine viral diarrhea virus from sika deer in China. Virol J.
2011;8:83.
59. Givens MD, Heath AM, Brock KV, et al. Detection of bovine
viral diarrhea virus in semen obtained after inoculation of serone-
gative postpubertal bulls. Am J Vet Res. 2003;64:428–434.
60. Givens MD, Marley MS. Immunology of chronic BVDV infec-
tions. Biologicals. 2013;41:26–30.
61. Glew EJ, Carr BV, Brackenbury LS, et al. Differential effects of
bovine viral diarrhoea virus on monocytes and dendritic cells.
J Gen Virol. 2003;84:1771–1780.
62. Glew EJ, Howard CJ. Antigen-presenting cells from calves persis-
tently infected with bovine viral diarrhoea virus, a member of the
Flaviviridae, are not compromised in their ability to present viral
antigen. J Gen Virol. 2001;82:1677–1685.
63. Goyal SM, Bouljihad M, Haugerud S, et al. Isolation of bovine
viral diarrhea virus from an alpaca. J Vet Diagn Invest. 2002;
14:523–525.
64. Greiser-Wilke I, Liebler E, Haas L, et al. Distribution of cyto-
pathogenic and noncytopathogenic bovine virus diarrhea virus
in tissues from a calf with experimentally induced mucosal dis-
ease using antigenic and genetic markers. Arch Virol Suppl.
1993;7:295–302.
65. Grooms DL, Brock KV, Pate JL, et al. Changes in ovarian folli-
cles following acute infection with bovine viral diarrhea virus.
Theriogenology. 1998;49:595–605.
66. Grooms DL, Brock KV, Ward LA. Detection of bovine viral diar-
rhea virus in the ovaries of cattle acutely infected with bovine
viral diarrhea virus. J Vet Diagn Invest. 1998;10:125–129.
67. Grummer B, Bendfeldt S, Wagner B, et al. Induction of the intrin-
sic apoptotic pathway in cells infected with cytopathic bovine
virus diarrhoea virus. Virus Res. 2002;90:143–153.
Downloaded from vet.sagepub.com at UNIV ARIZONA LIBRARY on June 15, 2014
461
68. Grummer B, Moennig V, Greiser-Wilke I. Cytopathogenic bovine
viral diarrhea viruses induce apoptosis in bovine cell cultures [in
German]. Dtsch Tierarztl Wochenschr. 1998;105:29–31.
69. Haines DM, Clark EG, Dubovi EJ. Monoclonal antibody-based
immunohistochemical detection of bovine viral diarrhea virus in
formalin-fixed, paraffin-embedded tissues. Vet Pathol. 1992;29:
27–32.
70. Hausmann Y, Roman-Sosa G, Thiel HJ, et al. Classical swine
fever virus glycoprotein E rns is an endoribonuclease with an
unusual base specificity. J Virol. 2004;78:5507–5512.
71. Heinz FX, Collet MS, Purcell RH, et al. Genus Pestivirus. New
York, NY: Academic Press; 2000.
72. Henningson JN, Steffen DJ, Topliff CL, et al. Systemic distribu-
tion of viral antigen in alpacas persistently infected with bovine
pestivirus. Vet Pathol. 2013;50:308–317.
73. Hessman BE, Sjeklocha DB, Fulton RW, et al. Acute bovine viral
diarrhea associated with extensive mucosal lesions, high morbid-
ity, and mortality in a commercial feedlot. J Vet Diagn Invest.
2012;24:397–404.
74. Houe H. Epidemiological features and economical importance of
bovine virus diarrhoea virus (BVDV) infections. Vet Microbiol.
1999;64:89–107.
75. Howard CJ, Clarke MC, Sopp P, et al. Immunity to bovine virus
diarrhoea virus in calves: the role of different T-cell subpopula-
tions analysed by specific depletion in vivo with monoclonal anti-
bodies. Vet Immunol Immunopathol. 1992;32:303–314.
76. Jewett CJ, Kelling CL, Frey ML, et al. Comparative pathogenicity
of selected bovine viral diarrhea virus isolates in gnotobiotic
lambs. Am J Vet Res. 1990;51:1640–1644.
77. Kelling CL, Hunsaker BD, Steffen DJ, et al. Characterization of
protection from systemic infection and disease by use of a
modified-live noncytopathic bovine viral diarrhea virus type 1
vaccine in experimentally infected calves. Am J Vet Res. 2005;
66:1785–1791.
78. Kelling CL, Hunsaker BD, Steffen DJ, et al. Characterization of
protection against systemic infection and disease from experi-
mental bovine viral diarrhea virus type 2 infection by use of a
modified-live noncytopathic type 1 vaccine in calves. Am J Vet
Res. 2007;68:788–796.
79. Kelling CL, Steffen DJ, Cooper VL, et al. Effect of infection with
bovine viral diarrhea virus alone, bovine rotavirus alone, or
concurrent infection with both on enteric disease in gnotobiotic
neonatal calves. Am J Vet Res. 2002;63:1179–1186.
80. Kelling CL, Steffen DJ, Topliff CL, et al. Comparative virulence
of isolates of bovine viral diarrhea virus type II in experimentally
inoculated six- to nine-month-old calves. Am J Vet Res. 2002;63:
1379–1384.
81. Kelling CL, Topliff CL. Bovine maternal, fetal and neonatal
responses to bovine viral diarrhea virus infections. Biologicals.
2013;41:20–25.
82. Kennedy JA, Mortimer RG, Powers B. Reverse transcription-
polymerase chain reaction on pooled samples to detect bovine
viral diarrhea virus by using fresh ear-notch-sample supernatants.
J Vet Diagn Invest. 2006;18:89–93.
83. Krametter-Froetscher R, Duenser M, Preyler B, et al. Pestivirus infec-
tion in sheep and goats in west Austria. Vet J. 2010;186:342–346.
462
84. Lackner T, Muller A, Pankraz A, et al. Temporal modulation of an
autoprotease is crucial for replication and pathogenicity of an
RNA virus. J Virol. 2004;78:10765–10775.
85. Larsson B. Failure to induce mucosal disease in cattle persistently
infected with bovine viral diarrhea virus by treatment with adreno-
corticotropic hormone. Acta Vet Scand. 1988;29:1–8.
86. Lee SR, Nanduri B, Pharr GT, et al. Bovine viral diarrhea virus
infection affects the expression of proteins related to professional
antigen presentation in bovine monocytes. Biochim Biophys Acta.
2009;1794:14–22.
87. Lee SR, Pharr GT, Boyd BL, et al. Bovine viral diarrhea
viruses modulate Toll-like receptors, cytokines and co-
stimulatory molecules genes expression in bovine peripheral
blood monocytes. Comp Immunol Microbiol Infect Dis.
2008;31:403–418.
88. Liebler-Tenorio EM, Kenklies S, Greiser-Wilke I, et al. Inci-
dence of BVDV1 and BVDV2 infections in cattle submitted for
necropsy in northern Germany. J Vet Med B Infect Dis Vet Pub-
lic Health. 2006;53:363–369.
89. Liebler-Tenorio EM, Ridpath JE, Neill JD. Distribution of viral
antigen and tissue lesions in persistent and acute infection with
the homologous strain of noncytopathic bovine viral diarrhea
virus. J Vet Diagn Invest. 2004;16:388–396.
90. Liebler-Tenorio EM, Ridpath JF, Neill JD. Distribution of viral
antigen and development of lesions after experimental infection
of calves with a BVDV 2 strain of low virulence. J Vet Diagn
Invest. 2003;15:221–232.
91. Liebler-Tenorio EM, Ridpath JF, Neill JD. Lesions and tis-
sue distribution of viral antigen in severe acute versus subcli-
nical acute infection with BVDV2. Biologicals. 2003;31:
119–122.
92. Liebler EM, Waschbusch J, Pohlenz JF, et al. Distribution of
antigen of noncytopathogenic and cytopathogenic bovine virus
diarrhea virus biotypes in the intestinal tract of calves following
experimental production of mucosal disease. Arch Virol Suppl.
1991;3:109–124.
93. Loy JD, Gander J, Mogler M, et al. Development and evalua-
tion of a replicon particle vaccine expressing the E2 glyco-
protein of bovine viral diarrhea virus (BVDV) in cattle.
Virol J. 2013;10:35.
94. Lunardi M, Headley SA, Lisboa JA, et al. Outbreak of acute
bovine viral diarrhea in Brazilian beef cattle: clinicopathological
findings and molecular characterization of a wild-type BVDV
strain subtype 1b. Res Vet Sci. 2008;85:599–604.
95. Malmquist WA. Bovine viral diarrhea-mucosal disease, etiol-
ogy, pathogenesis, and applied immunology. J Am Vet Med
Assoc. 1968;152:763–769.
96. Matzener P, Magkouras I, Rumenapf T, et al. The viral RNase
E(rns) prevents IFN type-I triggering by pestiviral single- and
double-stranded RNAs. Virus Res. 2009;140:15–23.
97. McClurkin AW, Bolin SR, Coria MF. Isolation of cytopathic and
noncytopathic bovine viral diarrhea virus from the spleen of cat-
tle acutely and chronically affected with bovine viral diarrhea. J
Am Vet Med Assoc. 1985;186:568–569.
98. McClurkin AW, Coria MF, Cutlip RC. Reproductive perfor-
mance of apparently healthy cattle persistently infected with
Downloaded from vet.sagepub.com at UNIV ARIZONA LIBRARY on June 15, 2014
Veterinary Pathology 51(2)
bovine viral diarrhea virus. J Am Vet Med Assoc. 1979;174:
1116–1119.
99. McClurkin AW, Littledike ET, Cutlip RC, et al. Production of
cattle immunotolerant to bovine viral diarrhea virus. Can J
Comp Med. 1984;48:156–161.
100. McGowan MR, Kirkland PD. Early reproductive loss due to
bovine pestivirus infection. Br Vet J. 1995;151:263–270.
101. Meyers G, Ege A, Fetzer C, et al. Bovine viral diarrhea virus:
prevention of persistent fetal infection by a combination of two
mutations affecting Erns RNase and Npro protease. J Virol.
2007;81:3327–3338.
102. Miller LC, Fox JM. Apoptosis and porcine reproductive and
respiratory syndrome virus. Vet Immunol Immunopathol. 2004;
102:131–142.
103. Montgomery DL. Distribution and cellular heterogeneity of
bovine viral diarrhea viral antigen expression in the brain of per-
sistently infected calves: a new perspective. Vet Pathol. 2007;44:
643–654.
104. Montgomery DL, Van Olphen A, Van Campen H, et al. The fetal
brain in bovine viral diarrhea virus–infected calves: lesions,
distribution, and cellular heterogeneity of viral antigen at 190
days gestation. Vet Pathol. 2008;45:288–296.
105. Munoz-Zanzi CA, Johnson WO, Thurmond MC, et al. Pooled-
sample testing as a herd-screening tool for detection of bovine
viral diarrhea virus persistently infected cattle. J Vet Diagn
Invest. 2000;12:195–203.
106. Nelson DD, Dark MJ, Bradway DS, et al. Evidence for persistent
bovine viral diarrhea virus infection in a captive mountain goat
(Oreamnos americanus). J Vet Diagn Invest. 2008;20:752–759.
107. Njaa BL, Clark EG, Janzen E, et al. Diagnosis of persistent
bovine viral diarrhea virus infection by immunohistochemical
staining of formalin-fixed skin biopsy specimens. J Vet Diagn
Invest. 2000;12:393–399.
108. Nobiron I, Thompson I, Brownlie J, et al. Cytokine adjuvancy of
BVDV DNA vaccine enhances both humoral and cellular
immune responses in mice. Vaccine. 2001;19:4226–4235.
109. Odeon AC, Kelling CL, Marshall DJ, et al. Experimental
infection of calves with bovine viral diarrhea virus genotype II
(NY-93). J Vet Diagn Invest. 1999;11:221–228.
110. Ohmann HB.Experimental fetal infection with bovine viral diar-
rhea virus, II: morphological reactions and distribution of viral
antigen. Can J Comp Med. 1982;46:363–369.
111. Ohmann HB. Pathogenesis of bovine viral diarrhoea-mucosal
disease: distribution and significance of BVDV antigen in
diseased calves. Res Vet Sci. 1983;34:5–10.
112. Olafson P, Mac CA, Fox FH. An apparently new transmissible
disease of cattle. Cornell Vet. 1946;36:205–213.
113. Palomares RA, Marley SM, Givens MD, et al. Bovine viral diar-
rhea virus fetal persistent infection after immunization with a
contaminated modified-live virus vaccine. Theriogenology.
2013;79:1184–1195.
114. Palomares RA, Walz HG, Brock KV. Expression of type I
interferon-induced antiviral state and pro-apoptosis markers
during experimental infection with low or high virulence
bovine viral diarrhea virus in beef calves. Virus Res. 2013;
173:260–269.
Brodersen
115. Passler T, Walz HL, Ditchkoff SS, et al. Distribution of
bovine viral diarrhoea virus antigen in persistently infected
white-tailed deer (Odocoileus virginianus). J Comp Pathol.
2012;147:533–541.
116. Pedrera M, Gomez-Villamandos JC, Risalde MA, et al. Charac-
terization of apoptosis pathways (intrinsic and extrinsic) in
lymphoid tissues of calves inoculated with non-cytopathic
bovine viral diarrhoea virus genotype-1. J Comp Pathol. 2012;
146:30–39.
117. Pedrera M, Sanchez-Cordon PJ, Romero-Trevejo JL, et al. Mor-
phological changes and virus distribution in the ileum of
colostrum-deprived calves inoculated with non-cytopathic
bovine viral diarrhoea virus genotype-1. J Comp Pathol. 2009;
141:52–62.
118. Perler L, Schweizer M, Jungi TW, et al. Bovine viral diar-
rhoea virus and bovine herpesvirus-1 prime uninfected
macrophages for lipopolysaccharide-triggered apoptosis by
interferon-dependent and -independent pathways. J Gen
Virol. 2000;81:881–887.
119. Peterhans E, Jungi TW, Schweizer M. BVDV and innate immu-
nity. Biologicals. 2003;31:107–112.
120. Pritchard WR. The bovine viral diarrhea-mucosal disease
complex. Adv Vet Sci Comp Med. 1963;41:1–47.
121. Ramsey FK. Pathology of a mucosal disease of cattle. Ames:
Iowa State College; 1956:32.
122. Raya AI, Gomez-Villamandos JC, Sanchez-Cordon PJ, et al.
Virus distribution and role of thymic macrophages during
experimental infection with noncytopathogenic bovine viral
diarrhea virus type 1. Vet Pathol. 2012;49:811–818.
123. Reitt K, Hilbe M, Voegtlin A, et al. Aetiology of bovine abortion
in Switzerland from 1986 to 1995—a retrospective study with
emphasis on detection of Neospora caninum and Toxoplasma
gondii by PCR. J Vet Med A Physiol Pathol Clin Med. 2007;
54:15–22.
124. Ridpath JF. Immunology of BVDV vaccines. Biologicals. 2013;
41:14–19.
125. Ridpath JF, Bolin SR. Delayed onset postvaccinal mucosal
disease as a result of genetic recombination between genotype
1 and genotype 2 BVDV. Virology. 1995;212:259–262.
126. Ridpath JF, Bolin SR, Dubovi EJ. Segregation of bovine viral
diarrhea virus into genotypes. Virology. 1994;205:66–74.
127. Ridpath JF, Driskell EA, Chase CC, et al. Reproductive tract
disease associated with inoculation of pregnant white-tailed deer
with bovine viral diarrhea virus. Am J Vet Res. 2008;69:
1630–1636.
128. Ridpath JF, Fulton RW, Kirkland PD, et al. Prevalence and anti-
genic differences observed between bovine viral diarrhea virus
subgenotypes isolated from cattle in Australia and feedlots in the
southwestern United States. J Vet Diagn Invest. 2010;22:
184–191.
129. Roth JA, Kaeberle ML. Suppression of neutrophil and lympho-
cyte function induced by a vaccinal strain of bovine viral diar-
rhea virus with and without the administration of ACTH. Am J
Vet Res. 1983;44:2366–2372.
130. Saliki JT, Huchzermeier R, Dubovi EJ. Evaluation of a new
sandwich ELISA kit that uses serum for detection of cattle
Downloaded from vet.sagepub.com at UNIV ARIZONA LIBRARY on June 15, 2014
463
persistently infected with BVD virus. Ann N Y Acad Sci. 2000;
916:358–363.
131. Scherer CF, Flores EF, Weiblen R, et al. Experimental infection
of pregnant ewes with bovine viral diarrhea virus type-2
(BVDV-2): effects on the pregnancy and fetus. Vet Microbiol.
2001;79:285–299.
132. Schneider R, Unger G, Stark R, et al. Identification of a struc-
tural glycoprotein of an RNA virus as a ribonuclease. Science.
1993;261:1169–1171.
133. Schweizer M, Matzener P, Pfaffen G, et al. ‘‘Self’’ and ‘‘non-
self’’ manipulation of interferon defense during persistent infec-
tion: bovine viral diarrhea virus resists alpha/beta interferon
without blocking antiviral activity against unrelated viruses
replicating in its host cells. J Virol. 2006;80:6926–6935.
134. Schweizer M, Peterhans E. Oxidative stress in cells infected with
bovine viral diarrhoea virus: a crucial step in the induction of
apoptosis. J Gen Virol. 1999;80(pt 5):1147–1155.
135. Scruggs DW, Fleming SA, Maslin WR, et al. Osteopetrosis, ane-
mia, thrombocytopenia, and marrow necrosis in beef calves
naturally infected with bovine virus diarrhea virus. J Vet Diagn
Invest. 1995;7:555–559.
136. Shimizu M.Current situation of bovine virus diarrhoea-
mucosal disease (BVD-MD) virus infections and their anti-
genic diversity in Hokkaido, Japan. Rev Sci Tech. 1990;9:
181–194.
137. Smirnova NP, Bielefeldt-Ohmann H, Van Campen H, et al.
Acute non-cytopathic bovine viral diarrhea virus infection
induces pronounced type I interferon response in pregnant cows
and fetuses. Virus Res. 2008;132:49–58.
138. Smirnova NP, Ptitsyn AA, Austin KJ, et al. Persistent fetal infec-
tion with bovine viral diarrhea virus differentially affects mater-
nal blood cell signal transduction pathways. Physiol Genomics.
2009;36:129–139.
139. Smirnova NP, Webb BT, Bielefeldt-Ohmann H, et al. Develop-
ment of fetal and placental innate immune responses during
establishment of persistent infection with bovine viral diarrhea
virus. Virus Res. 2012;167:329–336.
140. St-Louis MC, Massie B, Archambault D. The bovine viral diar-
rhea virus (BVDV) NS3 protein, when expressed alone in mam-
malian cells, induces apoptosis which correlates with caspase-8
and caspase-9 activation. Vet Res. 2005;36:213–227.
141. Stoffregen B, Bolin SR, Ridpath JF, et al. Morphologic lesions in
type 2 BVDV infections experimentally induced by strain
BVDV2-1373 recovered from a field case. Vet Microbiol.
2000;77:157–162.
142. Thompson K. Bones and joints. In: Maxie MG, ed. Jubb, Kennedy
and Palmer’s Pathology of Domestic Animals. 5th ed. Vol 1. Phi-
ladelphia, PA: Saunders Elsevier; 2007:66–67.
143. Thur B, Zlinszky K, Ehrensperger F. Immunohistochemical
detection of bovine viral diarrhea virus in skin biopsies: a reli-
able and fast diagnostic tool. Zentralbl Veterinarmed B. 1996;
43:163–166.
144. Tyler DE, Ramsey FK. Comparative pathologic immunologic
and clinical responses produced by selected agents of bovine
mucosal disease—virus diarrhea complex. Am J Vet Res. 1965;
26:903–913.
464
145. Uttenthal A, Hoyer MJ, Grondahl C, et al. Vertical transmission
of bovine viral diarrhoea virus (BVDV) in mousedeer (Tragulus
javanicus) and spread to domestic cattle. Arch Virol. 2006;151:
2377–2387.
146. Vassilev VB, Donis RO. Bovine viral diarrhea virus induced
apoptosis correlates with increased intracellular viral RNA accu-
mulation. Virus Res. 2000;69:95–107.
147. Vega S, Rosell R, Paton DJ, et al. Antigenic characterization of
bovine viral diarrhoea virus isolates from Spain with a panel of
monoclonal antibodies. J Vet Med B Infect Dis Vet Public
Health. 2000;47:701–706.
148. Vilcek S, Durkovic B, Kolesarova M, et al. Genetic diversity of
BVDV: consequences for classification and molecular epide-
miology. Prev Vet Med. 2005;72:31–35; discussion 215–219.
149. Webb BT, Norrdin RW, Smirnova NP, et al. Bovine viral diarrhea
virus cyclically impairs long bone trabecular modeling in experi-
mental persistently infected fetuses. Vet Pathol. 2012;49:930–940.
Downloaded from vet.sagepub.com at UNIV ARIZONA LIBRARY on June 15, 2014
Veterinary Pathology 51(2)
150. Weber F, Wagner V, Rasmussen SB, et al. Double-stranded
RNA is produced by positive-strand RNA viruses and DNA
viruses but not in detectable amounts by negative-strand RNA
viruses. J Virol. 2006;80:5059–5064.
151. Wilcock BP. Eye and Ear. In: Maxie MG, ed. Jubb, Kennedy, and
Palmer’s Pathology of Domestic Animals. 5th ed. Vol 1. Philadel-
phia, PA: Saunders Elsevier; 2007:477
152. Wilhelmsen CL, Bolin SR, Ridpath JF, et al. Lesions and localiza-
tion of viral antigen in tissues of cattle with experimentally induced
or naturally acquired mucosal disease, or with naturally acquired
chronic bovine viral diarrhea. Am J Vet Res. 1991;52:269–275.
153. Wittum TE, Grotelueschen DM, Brock KV, et al. Persistent
bovine viral diarrhoea virus infection in US beef herds. Prev Vet
Med. 2001;49:83–94.
154. Zhang G, Aldridge S, Clarke MC, et al. Cell death induced by
cytopathic bovine viral diarrhoea virus is mediated by apoptosis.
J Gen Virol. 1996;77(pt 8):1677–1681.