Journal of Reproduction and Development, Vol.

56, Suppl, 2010

Infectious Causes of Reproductive Disorders in Cattle

Han Sang YOO
Department of Infectious Diseases, BK21 for Veterinary Science and KRF Priority Zoonotic Research Institute, College of
Veterinary Medicine, Seoul National University, Seoul 151-742, Korea
Abstract. The incidences of reproductive disorders in bovine are increasing over years. This scenario is further
aggravating due to more emphasis on selection and rearing of animal for specific commercial purposes which
compromises livestock reproduction. Reproductive disorders like infertility and abortions in cattle are major problems
in the bovine industry. The reproductive disorders might be caused by several different agents such as physical agents,
chemical agents, biological agents, etc. Also, the causative agent and pathogenesis of reproductive disorders are
influenced by various factors including environmental factor. The exact causes may not be evident and are often
complicated with multiple causative agents. Thus, there is a need for multi-faceted approach to understand correlation
of various factors with reproductive performance. Of the agents, infectious biological agents are significant cause of
reproductive disorder and are of high priority in the bovine industry. These factors are not only related to the
prosperity of bovine industry but are also important from public health point of view because of their zoonotic
potentials. Several infectious agents like bacterial, viral, protozoon, chlamydial and fungal agents are known to have
direct impact on reproductive health of cattle. These diseases can be arranged and discussed in different groups based
on the causative agents.
Key words: Cattle, Infectious agents, Reproductive disorders
(J. Reprod. Dev. 56: S53–S60, 2010)

eproductive efficacy is one of the main factors that make the
cattle industry economically and clinically sound. Factors that
affect reproduction have a substantial economic impact on produc-
ers who want the production of viable and healthy offspring [1–3].
Causes of reproductive disorders such as infertility and abortion are
numerous and range from simple management errors to compli-
cated multi-factorial disease complexes due to several different
factors such as physical, chemical, biological, and environmental
agents [1–3]. Although there have been attempts to improve the
reproductive efficacy of bovine, the incidences of reproductive dis-
orders in bovine have still been increasing over the years. The
exact cause of this may not be evident and is often complicated by
multiple causative agents. Thus, there is a need for a multi-faceted
approach to understanding the correlation of various factors with
reproductive performance in the bovine industry.
Of the agents, infectious biological agents are significant causes
of reproductive disorders and are being given high priority in the
bovine industry [1–5]. These factors not only have a significant
impact on the prosperity of the bovine industry but are also impor-
tant from the public health point of view because of their zoonotic
potentials [2–4]. Several infectious agents such as bacterial, viral,
protozoal, and other agents such as chlamydial and fungal agents
are known to have a direct impact on the reproductive health of cat-
tle. The causes of reproductive failure will be discussed under
different categories based on the causative agents.
Bacterial Agents
Of the agents, pathogenic bacteria have been receiving increased
Correspondence: HS Yoo (e-mail: yoohs@snu.ac.kr)
attention as important causes of reproductive diseases, including
abortion and sterility, in cattle. Brucella abortus, Leptospira inter-
rogans, Camphlyobacter fetus, Listeria monocytogenesis, and
Salmonella spp. have been implicated in reproductive failure in
bovine. These bacterial agents and how they cause reproductive
failure in bovine are discussed next.
Brucella abortus
Bovine brucellosis caused by Brucella abortus results in abor-
tions in female cattle and infertility in male cattle that are infected
and discharge bacteria in their semen [4–6]. Bovine brucellosis is
widely seen in most parts of the world except for some countries
such as Australia, Japan, and northern Europe (OIE, 2005). The
importance of the disease is related not only to economic losses in
animal production, but also to risks to human health [6–8]. The
causative bacterium is a Gram-negative coccobacilli that is non-
spore-forming, non-motile, and non-encapsulated. Nine species of
Brucella have been identified within the genus: B. abortus, B.
canis, B. melitensis, B. neotomae, B. ovis, B. suis, B. ceti, B. pinni-
pedia and B. microti [6, 7]. Although most of these species have a
wide range of hosts, B. abortus is the main causative agent of bru-
cellosis in bovine [4, 6]. Infection with B. abortus showed the long
and variable incubation period and the occurrence of latent infec-
tion in calves born to infected cows without serological response
upon future infection [5, 7, 8]. Also, the bacteria can survive inside
the cells of the infected host by creating a membrane-bound com-
partment that prevents their fusion with lysosomes and supports
intracellular replication [7, 9]. In addition, brucella is easily aero-
solized, highly invasive, and extremely infectious. Such factors
make it difficult to eradicate bovine brucellosis. The disease is
marked by late abortion with significant adverse effects on the male
 S54 YOO
reproductive system, such as conception failure, epididmymitis,
and orchitis [4, 6].
The most common infection route is the digestive tract, through
the ingestion of contaminated feeds or water with infected placen-
tas or aborted fetuses. After the ingestion, the bacteria are
internalized by the M cells in the Peyer’s patches [4]. They then
spread to the regional lymph nodes where, as facultative intracellu-
lar microorganisms, they proliferate within macrophages.
Subsequently, they spread via the blood stream to other tissues,
particularly the pregnant uterus [4, 6].
In cattle, B. abortus has a marked tropism for placental tissues,
which results in gross lesions particularly in placentomes, which
become friable, covered with fibrinous exudates, and fetid [4, 6].
Usually, there are no gross lesions in the mammary glands even
though the bacteria are secreted in the milk [4, 6]. Aborted fetuses
show variable degrees of autolysis and may be oedematous; and in
some cases, the abdominal organs may be covered with fibrin [4,
6]. Pneumonia has been described as the most common lesion in
fetuses that had been aborted due to B. abortus infection [4, 10].
The diagnosis of brucellosis is based on direct or indirect labora-
tory methods. Direct methods such as the isolation and
identification of the bacterial agent have high specificity, but are
time-consuming and require facilities with an appropriate level of
biosafety. Therefore, PCRs, including multiplex PCR, were
recently developed to detect the presence of the genetic materials of
the bacteria [8, 11]. Serological methods such as tube agglutination
and slide agglutination are used more often because they are
quicker and less costly. Also, immunofluorescence and immuno-
histochemical methods as alternative methods of detecting B.
abortus in tissues have been using [4]. Immunofluorescence has
high specificity but low sensitivity [4]. Immunohistochemical
examination of paraffin-wax-embedded tissues to detect the pres-
ence of B. abortus antigens is not only both sensitive and specific,
but it also clearly shows the tissue morphology. It can thus demon-
strate the distribution of organisms in the tissues, which is a
valuable attribute in the study of the pathogenesis of B. abortus
infection [4, 12]. Bacterial isolation, clinical symptoms and sero-
logical tests are also used for the diagnosis. Among the serological
tests, the standard tube agglutination test is recognized as the stan-
dard method for the diagnosis of bovine brucellosis in OIE.
Several vaccines, including inactivated and live attenuated vac-
cines, against bovine brucellosis have been used in many regions of
the world for the prevention and control of the disease [5–7]. How-
ever, the use of vaccine differs depending on the prevalence of the
disease and the eradication program in the countries.
Leptospira interrogans
Leptospirosis is an economically important bacterial infection of
livestock that causes not only reproductive losses due to abortions,
stillbirths, and infertility but also non-reproductive losses due to
septicemia and nephritis. Leptospirosis is also known as the most
widespread zoonotic disease in the world [1, 2, 13]. It is caused by
infection with a spirochete Leptospira. Based on genetic
sequences, there are currently more than 15 recognized genome
species of Leptospira, seven of which contain organisms that are
pathogenic to cattle [2, 14]. Approximately 200 different serovars
of pathogenic Leptospira have been identified throughout the world
[1, 2, 13]. The predominant serovar in cattle varies from one geo-
graphic region to another. In particular regions, different
Leptospira serovars are prevalent and are associated with one or
more maintenance hosts, which serve as reservoirs of the infection
[2, 13]. The most common cause of leptospirosis among cattle is
infection with Leptospira that belong to the serovar hardjo group.
Cattle appear to be the primary maintenance hosts of this serovar.
Other common causes of leptospirosis include the serovars pomona
and grippotyphosa in cattle [1, 13].
A recent estimate of Leptospira infection in a sample of U.S.
dairies and beef cow-calf operations is the overall herd prevalence
infection was approximately 35–50%, with most of the infections
likely due to serovar hardjo [2]. However, the prevalence and gen-
otypes of Leptospira differ depending on the areas. Typical
leptospiral infection occurs through bacterial exposure to mucous
membranes and generally results in no or relatively mild acute clin-
ical signs. As the consequence of serovar hardjo infection,
abortions, stillbirths, or birth of weak calves may occur, but the
symptoms are generally seen only when a cow is infected at the
first time of her pregnancy [3, 15]. An abortion may occur several
weeks after the infection of the dam without any association with
any obvious illness in the cow [2, 13].
After primary bacteremia, the Leptospira localize and persist
primarily in the kidneys and the genital tract. The Leptospira
located in the kidneys are voided in the urine and serve as sources
of infection to other animals [2, 8]. Persistent infection of the male
and female bovine genital tracts is a prominent feature of serovar
hardjo infections and may last for more than 12 months [2, 16].
Persistent infection of the reproductive tract may be the most eco-
nomically important manifestation of serovar hardjo infection.
Infertility also is associated with this infection.
Diagnostic tests of leptospira infection include fluorescent anti-
body testing (FAT), culture of bacteria, PCR and silver staining and
immune-histochemistry with tissue samples. Also, serological
tests such as ELISA, microscopic agglutination testing (MAT) are
established for the diagnosis of leptospiral infection [1, 8, 13].
Campylobacter fetus
Bovine genital campylobacteriosis (BGC) is a bacterial venereal
disease that may lead to infertility, early embryonic mortality, and
abortion [12, 17–19]. BGC occurs worldwide and causes high eco-
nomic losses to the bovine industry. The disease is transmitted
through sexual contact among bovine. Thus, it has been known as
bovine veneral campylobacteriosis because of the transmission
route. The incidence of BCG has been reduced in many industrial-
ized countries due to the introduction of artificial insemination (AI)
in combination with veterinary health control programs and the
strong host specificity of this bacterium [17, 20]. However, BGC is
still a major problem of the cattle industry in developing countries
because of non-optimal health surveillance programs, including the
introduction of AI [17]. There are two subspecies of Campylo-
bacter fetus: C. fetus subsp. fetus and C. fetus subsp. venerealis. C.
fetus subsp. venerealis adapts well to the bovine reproductive tract,
whereas C. fetus subsp. fetus is found in the intestine and causes
abortion in sheep and cattle [12, 17–19]. Both subspecies charac-
 INFECTIOUS CAUSES OF REPRODUCTIVE DISORDERS IN CATTLES
teristically produce comma-shaped or S-shaped Gram-negative
bodies and microaerophilites [19, 20]. Therefore, the optimal
growth conditions of the organisms require an atmosphere of 10–
20% CO2, and below 5% oxygen. The optimal temperature for
their growth is 37 C. Although the bacteria can grow on serum,
blood agar, and thiol agar, antibiotics such as novobiocin and baci-
tracin can be introduced to them to inhibit the growth of
contaminants [20].
C. fetus subsp. venerealis is an obligate parasite of the bovine
genitalia. The infection is unapparent and involves the epithelium
of the penis and the fornix of the prepuce of bulls [18, 19]. In a sus-
ceptible herd, the introduction of the organism is followed by a
period of infertility of all the breeding females (120 days). How-
ever, only newly introduced susceptible animals show signs of
infertility in chronically infected herds [18, 20]. The organism
localizes in the anterior vagina and cervix during the ovulatory
phase, but does not invade the uterus and oviducts until the proges-
tational phase. Moderate endometritis and salpingitis then result
and persist for several weeks to a few months [18, 20]. Animals
usually regain fertility within five months after the elimination of
the infection from the uterus and the oviducts. The antibodies in
the vaginal secretions are predominantly in the IgA class, and they
immobilize the organism so that it could not penetrate higher into
the tract. Although vaccines have been introduced to prevent and
terminate natural infections in bulls, C. fetus subsp. venerealis
infection in cattle herds that breed naturally is self-limiting because
of the normal immune response in infected animals [20]. The
infection persists, however, in some cows and bulls. Thus, they can
be sources of infection of susceptible virgin heifers and newly
introduced stock [20]. C. fetus subsp. fetus occasionally causes
abortion, however, during the latter half of the gestation phase of
cattle [18, 20]. The pathogenesis of the infection is similar to that
of C. fetus subsp. venerealis, but its infection route is different.
Infection with C. fetus subsp. fetus is probably caused by the inges-
tion of infectious materials rather than by venereal transmission
[18, 20]. The organism localizes in the placentomes after a period
of bacteremia, after which placentitis develops and abortion occurs
toward the end of the gestation period. Only heavily gravid ani-
mals are susceptible to bacterial invasion of the placenta with
subsequent abortion. The incubation period is seven to 25 days.
Animals usually have a vaginal discharge for several days before
they experience abortion. Vaccination has been shown to be effec-
tive in the prevention of C. fetus subsp. fetus abortion in sheep
flocks. This bacterium could be diagnosed through (i) direct-
impression smearing of aborted materials such as cotyledons and
fetal stomach contents, (ii) isolation and identification of the bacte-
ria, and (iii) detection of agglutinating antibodies in the cervical
mucus [17, 18, 20].
Listeria monocytogenes
Listeria monocytogenes are Gram-positive, non-acid-fast organ-
isms that appear as 1–2 μm-long small rods. L. monocytogenes are
non-spore-forming and facultative intracellular bacteria that grow
on most bacterial media. They produce two different antigens:
somatic (O) and flagellar (H) antigens. Fifteen O antigens (I–XV)
and five H antigens (A–E) have been identified [21]. L. monocyto-
S55
genes are widely distributed in nature and in animal and human
feces. They can survive for years in soil, milk, silage, and feces
even though their survival time outside their host varies depending
on their environment [21]. Although most animals are exposed to
L. monocytogenes in their lifetime due to the wide distribution of
the organisms in the environment, predisposing factors such as
intercurrent diseases, climatic and other stresses, and pregnancy are
closely related to the occurrence of the disease [21, 22]. Predispos-
ing factors that diminish the host’s cell-mediated immune
competence may allow the parasite to escape from phagocytic cells
and cause disease due to out of multiplication of the bacteria. The
most of outbreak in cattle may occur in winter and early spring.
Although the initial mode of entry of L. monocytogenes into an ani-
mal is usually via ingestion, the organism can localize in the
placentomes and enter the amniotic fluid in pregnant animals and
multiplies [21, 22]. Abortions in cattle usually occur in the second
half of their pregnancy [21]. The antibodies do not play an impor-
tant role in the resolution of the disease because immunity to L.
monocytogenes is cell-mediated by means of rapidly dividing,
short-lived T lymphocytes [21]. Routine diagnosis tests such as by
isolating the organism and the clinical symptoms, and appropriate
therapeutic and preventive measures are helpful.
Salmonella spp.
Salmonella enterica subsp. enterica serovar Dublin causes
infections in cattle, usually with serious clinical diseases [23, 24].
It is difficult to control the spread of the serovar Dublin in infected
farms partly due to the ability of Salmonella to survive in the envi-
ronment and certainly due to the presence of carrier animals.
Although infection with these bacteria in cattle is seen in several
clinical signs, abortion is one of its most common consequences
[23, 24]. Usually, other clinical signs of the disease, such as fever,
may precede the occurrence of abortion. The organism apparently
enters the fetus from the circulation of materials via the placenta,
and causes septicemia and death of the fetus [23, 24]. So far, there
has been no evidence of venereal transmission of the organism.
Edema of the subcutis and serosanguineous fluids in the peritoneal
cavity can be observed in the aborted fetus. Abortion occurs usu-
ally about 200 days after conception [23, 24]. Several vaccines
including live attenuated or killed vaccines have been used to pre-
vent S. dublin infection of cattle [23, 24].
In addition to the aforementioned bacterial agents, other acute
bacterial infectious diseases such as anthrax, black leg, Haemophi-
lus somonus infection might induce abortion in pregnant cows.
Viral Agents
Viral infections are also associated with reproductive disorders
and death among cattle. The Akabane virus, the Chuzan virus, and
the Aino virus are the major causes of abortion, premature births,
and stillbirths among pregnant cows. Deformed or infirm neonatal
calves have also been observed in herds that have been infected
with these viruses. Bovine herpes virus-1 and the bovine viral diar-
rhea virus can also induce reproductive failures with other clinical
signs.
 S56 YOO
Akabane virus
Akabane virus, an arbovirus, causes infection in ruminants and
is a disease of the bovine fetus. The Akabane virus is a member of
the genus Bunyaviridae. It falls under the Simbu group, which now
includes four other distinct viruses that also infect the biting midge
Culicoides brevitarsis and livestock [25, 26]. This virus causes
abortions in cattle and congenital abnormalities in calves born of
infected cows. In infected herds, stillborn or premature fetuses, as
well as deformed or infirm neonatal calves that die within a few
days after birth, have been observed. Non-purulent encephalomy-
elitis, characterized by necrosis of the nerve tissue and endothelial
proliferation, in an undifferentiated central nervous system was
observed in a three-month old fetus [27]. Polymyositis character-
ized by parenchymal degeneration and cellular infiltrates has been
observed in the skeletal muscle. The disease can be reproduced
experimentally in cattle and goat fetuses via placental infection and
intracerebral inoculation of calves. Experimental infection of bulls
caused a viremia that persisted for two to nine days, and neutraliz-
ing antibodies were detected within seven to ten days even though
the bulls did not show any clinical signs. The virus was not isolated
in the semen of the infected bulls, nor did the infection reduce the
quality of the bulls’ semen. The virus replicates in cell-culture sys-
tems such as in baby hamsters’ kidney cell line, and produce
cytopathic alteration. The virus replicates in hen’s eggs and causes
death and deformities of chick embryos. The virus can be detected
via PCR, even though it can be diagnosed based on the clinical
signs of the aborted fetus [27].
Chuzan virus
Chuzan virus, a member of the Palyam serogroup of orbiviruses
in the family Reoviridae, was isolated from the biting midge Culi-
coides oxytoma and from sentinel calves in 1985 [28, 29]. The
genome of this virus consists of 10 dsRNA segments that encode
seven structural (VP1-VP7) and four non-structural (NS1, NS2,
NS3, and NS3A) proteins. The virus was subsequently implicated
in an episoed of congenital abnormalities with the hydranenceph-
aly-cerebellar hypoplasia syndrome among calves in the Kyushu
district, from autumn 1985 through spring 1986 [28]. This virus
has been associated with various haematophagous arthropod vec-
tors and large mammals, principally cattle, in many parts of the
world, including Asia, Australia, and Africa [26, 29, 30]. Although
the virus seems to be involved in the occurrence of abortion and
congenital malformations in cattle, its pathogenic importance
remains unknown.
Aino virus
The Aino virus is an arthropod-borne virus that belongs to the
Orthobunyavirus genus of the Bunyaviridae family. Viruses of the
genus Orthobunyavirus are widely distributed in tropical areas of
the world, and some of them have been shown to induce symptom-
atic or asymptomatic illnesses in animals [26, 31]. The teratogenic
potential of the Aino virus in cattle, which are their natural verte-
brate hosts, has been documented in several regions. Serological
evidences from field investigations have indicated that a fetus is
infected with the virus mainly from 120 to 180 days of its gestation.
Data from several studies suggest that calves were not infected with
the virus in the uterus [30]. Also, the rate of transplacental infec-
tion of AINOV may be lower than that of AKAV, which may
explain the lower incidence of diseases caused by AINOV than by
AKAV in the field. Serological diagnosis and vaccines have been
developed to control the disease. The Bluetongue virus, which
belongs to the Reoviridae family, is another arthropod-borne virus
(arbovirus) that may cause abortion and/or congenital disorders
among calves.
Bovine herpesvirus 1
Bovine herpesvirus 1 (BHV-1), also known as the infectious
bovine rhinotracheitis (IBR) virus or the infectious pustular vul-
vovaginitis (IPV) virus, is a double-stranded DNA virus in the
Herpesviridae family. BHV-1 is primarily associated with several
clinical syndromes such as rhinotracheitis, pustular vulvovaginitis,
balanoposthitis, abortion, infertility, conjunctivitis, and encephali-
tis in bovine species [32–36]. The main sources of BHV-1
infection are nasal exudates and respiratory droplets, genital secre-
tions, semen, fetal fluids, and tissues [32–36]. Acute IPV usually
develops one to three days after mating, and the characteristic ini-
tial signs of IPV infection are frequent micturition and tail
swishing. Fever, depression, and anorexia are the most common
clinical signs among affected animals. They seek to avoid contact
of their tail with the vulva. The vulva is swollen and hyperemic,
with small pustules. Varying amounts of pus may be discharged
during secondary bacterial infection. The duration of the clinical
illness varies among individual animals. Generally, the lesions
heal 10–14 days after the onset of the disease, even though a puru-
lent vaginal discharge persists for several weeks in some animals.
Semen contaminated with BHV-1 can cause IPV, cervicitis with
copious mucopurulent discharge, and endometritis in cows, and
epididymitis in bulls [34, 35, 37, 38]. BHV-1 is also associated
with poor semen quality, which may result in a lower conception
rate. Abortions due to BHV-1 have been reported worldwide [32,
39, 40]. Abortions usually occur between four to seven months
during gestation, after natural infection or vaccination. The abor-
tion is due to the death of the fetus. Traction of the placenta may be
required because of the retained placenta. The cotyledons are usu-
ally blanched and degenerate without any gross lesions on the
aborted fetus. Endometritis, a poor conception rate, and a short
estrus could occur through insemination of infected semen [32, 33,
38, 41].
The BHV-1 virus becomes latent following primary infection
with a field isolate or vaccination with an attenuated strain [42] The
viral genomic DNA is usually detected in the sensory ganglia of the
trigeminal nerve in IBR and in the sacral spinal genitalia in IPV/
IPB cases [42, 43]. Latency may also occur in tonsillar lymphoid
cells and peripheral blood lymphocytes [42, 44]. Latent infected
virus may be reactivated by several stress factors such as transpor-
tation and climatic changes. Attenuated vaccine strains can remain
latent in the body, and vaccination does not provide protection
against the occurrence of a latent infection with a wild strain and
re-excretion of a wild strain [44, 45]. This virus causes late-term
abortions and conception failure from venereal diseases. Diagnosis
of this viral infection may be performed on the basis of serologic
tests that detect the virus or its components, or antibodies, and
 INFECTIOUS CAUSES OF REPRODUCTIVE DISORDERS IN CATTLES
through nucleic-acid-based tests that detect the genomic DNA via
PCR and nucleic acid hybridization and sequencing [46]. Isolation
of BHV-1 virus can be carried out using cell culture of primary or
secondary bovine kidneys, lungs, testes, turbinate, or tracheas, and
of established cell lines such as MDBK cells. Nasal swabs, con-
junctival swabs, vaginal swabs, preputial washing, placental
cotyledons of aborted fetuses, fetal livers, lungs, spleens, kidneys,
lymph nodes, mucous membranes of respiratory tracts, tonsils, and
lungs can be used to isolate the virus. Intranuclear viral inclusions
of Cowdry type A can be occasionally identified in the epithelial
cells of vaginal biopsy tissues collected in the early stage of IPV
and in tissues of aborted fetuses [47]. Electron microscopy may
also be used to identify the virus particles in clinical materials so as
to rapidly diagnose the virus, even though all herpes viruses are
indistinguishable. As an alternative method to detect the virus,
PCR which is more sensitive and faster than virus isolation, is
developed and applied to detect the BHV-1 virus [48]. However
the biggest concern is the how to avoid the false positive and nega-
tive. Therefore, Real-time PCR with satisfactory reproducibility
and high specificity and sensitivity was recently developed. Its sig-
nificant reduction of the time needed to detect amplified products
makes it a valuable alternative to the time- and labor-consuming
virus isolation method of detecting BHV-1 in extended semen [48,
49]. A battery of serological tests is available for the detection of
antibodies and rises in titers between the acute and convalescent
phases of the infection [34, 40].
Four kinds of vaccines are available to prevent this virus infec-
tion: modified live virus (MLV) vaccines, inactivated vaccines,
subunit vaccines, and marker vaccines [33, 34, 50]. MLV vaccines
are available for parental use and for the intranasal route. They can
induce a rapid immune response and long-lasting immunity, and
result in local and mucosal immunity. However, MLV vaccine
cannot use in non-immune pregnant cattle because of the possibil-
ity of abortion and latency of the disease. Inactivated vaccines
have been developed to overcome the disadvantages of MLV vac-
cines. They are safe for pregnant animals, stable in storage, and do
not cause shedding of the virus, and they do not cause immunosup-
pression, abortion, or latency, although they cannot prevent the
development of latency following exposure to the field virus. The
efficacy of inactivated vaccines might be lower, however, than that
of MLV vaccines. The subunit vaccine contains one or more anti-
gens of the virus that induce protective immunity and do not
contain nucleic acid and other components that might cause
unwanted side-effects. The marker vaccine is based on changes in
one or more microbial proteins, which allow the differentiation of
the vaccinated animals from the infected animals. The live marker
vaccine is safe for breeding cows, bulls, and pregnant cows. It is
also effective in the presence of maternal antibodies [33]. In sev-
eral countries such as the U.S., Australia, Korea, and European
countries, multivalent vaccines that contain parainfluenza type-3
(PI-3), BRSV, and the bovine viral diarrhea (BVD) virus are most
frequently used to control the infection.
Bovine viral diarrhea virus
Bovine viral diarrhea virus (BVDV) is a small, enveloped, posi-
tive-strand RNA virus with an approximately 12.3kb genome size
S57
in the Flaviviridae family and the genus Pestivirus, which also
includes the classical swine fever virus (CSFV) and the border dis-
ease virus of sheep [51–54]. BVDV is divided into two species
based on their 5’-UTRs: BVDV-1 and -2 [55, 56]. BVDV-1 strains
are particularly diverse, with at least 13 genetically and antigeneti-
cally different subgroups [53, 55, 57, 58]. Regardless of their
diversity, all BVD viruses are either of the cytopathic (cp) or the
non-cytopathic (ncp) biotype, based on the effect of the viral
growth in cultured cells [52, 53, 58]. The two biotypes also differ
with respect to their effects on infected animals. Only the ncp bio-
type can cause established a persistent infection of the fetus if the
dam is transiently infected between approximately 40 and 120 days
of its development [2, 51]. The clinical consequences of infection
with the BVD virus are as diverse as the genetic and antigenic
properties of the virus [57]. Diseases can result from different
pathogenetic mechanisms, depending on the different types of
infection. Most transient infections may be unapparent or mild,
and associated with low-grade fever, diarrhea, and coughing [51].
Rarely, acutely infected animals may suffer from high-grade fever
and bleeding in internal organs.
Reproductive losses associated with bovine viral diarrhea virus
(BVDV) infection were described in his first clinical description of
bovine viral diarrhea (BVD) by Olafson [59, 60]. Pregnant cows
subclinically infected with BVDV mostly experienced abortion
10–90 days later [2]. Today, reproductive losses may be the most
economically important consequence of BVDV infection, and there
are indications that the incidence of BVDV-related reproductive
losses is increasing in several countries, including the U.S [49].
BVDV uses the reproductive system to maintain and spread itself
in the cattle population by inducing immunotolerance after fetal
infection, and results in the birth of calves that are persistently
infected with the virus. Cattle that are persistently infected with
BVDV become major sources of the virus both within and among
farms. The reproductive losses from BVDV have numerous vari-
able clinical manifestations that range from an insidious reduction
in the reproductive performance at the herd level to devastating
abortion storms. The clinical features of BVDV infection differ
depending on the stage of the infection during the gestation phase,
and include early embryonic death, abortion, immunotolerance,
congenital defects, and congenital infection (normal/abnormal).
Persistent infection (PI) results when the susceptible cow or heifer
is exposed to the NCP strain of BVDV between 42 and 125 days of
gestation. The PI calves are born alive, become immunotolerant to
the infecting strain, and shed the virus throughout their lives [32,
61–63]. Therefore, such cattle are likely the most important reser-
voirs of the virus for susceptible cattle. The identification and
removal of PI animals are essential in the prevention of the spread
of BVDV, because they can serve as the natural reservoirs of the
virus [61, 62]. Thus, numerous methods have been developed to
diagnose both persistent infection and acute or transient infection
with BVDV. These include antigen-capture (AC) enzyme-linked
immunosorbent assay (ELISA), immunohistochemical (IHC) test-
ing, gel-based reverse-transcription (RT), real-time polymerase
chain reaction (PCR), and virus isolation using WBC lysates, tis-
sues, or whole blood in cell culture [64, 65]. Also, there are
numerous serological methods of measuring seroconversion from
 S58
acute infection, including ELISA and viral neutralization tests in
cell culture. Samples of milk, serum, and supernatants from skin
biopsies, usually ear notches, are used for the highly sensitive RT-
PCR, which detects viral RNA and is becoming a popular method
of detecting PI cattle. The absence of PI animals should lower the
incidence of the disease. Therefore, BVDV control requires proper
biosecurity to prevent additions of PI animals, optimal BVDV vac-
cination to maximize immunity, and continued vigilance in
identifying and removing PI cattle. Different strategies to eradicate
the disease are used depending on the country and the status of the
infection [66]. Several vaccines, including modified live, inacti-
vated, and combined vaccines, have also recently been developed
to control the disease. Modified live vaccines are considered most
effective because they produce immunogenic proteins through viral
replication, and offer long-lasting protection; but their safety is
questionable especially during pregnancy or in animals that have
contact with pregnant ones [66]. Inactivated vaccines are safe, but
they have disadvantages with respect to fetal protection, the dura-
tion of the protection, and production cost. The use of genetic
engineering methods is also being explored for the development of
new vaccines that may overcome the disadvantages of both vac-
cines—such as vector vaccines, DNA vaccines, subunit vaccines,
and marker or deletion mutants.
Other viruses such as Ibaraki virus, a family of Reoviridae, are
also being implicated in reproductive failures, including abortions,
in bovine [67].
Protozoal and Other Agents
Trichomoniasis caused by Tritrichomonas fetus and neosporosis
caused by Neospora caninum are the major pathogenic protozoans
that cause sterility and abortion, followed by other agents.
Tritrichomonas fetus
Tritrichomonas fetus is a sexually transmitted obligate proto-
zoan of cattle and is responsible for occult pregnancy losses in
farms where natural servicing of cattle is practiced. Tritrichomo-
nas fetus is found in the vagina, uterus, macerated fetus, prepuce,
penis, epididymis, and vas deferens. The organism has a consider-
able population of pleomorphisms, varies from 10 to 25 μm in
length, and has three anterior flagella and a long, trailing flagellum
that extends beyond the undulating membrane [68–70]. Although
three serotypes have been identified, antigenic types appear to have
no important role in the immunity of T. fetus because heifers immu-
nized with one serotype developed resistance to infection from
homologous and heterologous strains [70]. Bovine genital tri-
chomoniasis is a venereal disease that manifests in cows and
heifers in the form of infertility, abortion at up to five months after
breeding, pyometra, and occasional fetal mummification [70]. An
infected bull is usually responsible for spreading trichomoniasis in
a herd, and artificial insemination is recommended as a control
measure, when feasible. In male cattle, T. fetus is localized in the
secretions of the epithelial lining of the penis, prepuce, and distal
urethra. The most important feature of T. fetus infection of bulls is
the development of the chronic carrier state. No clear breed associ-
ation to bull susceptibility to trichomonosis has been established
YOO
yet. T. fetus infection has also been associated with bull factors
related to herd and herd management practice. In female cattle, T.
fetus gains entry through coital exposure to a persistently infected
male or through passive transfer from a previously uninfected bull
that has had recent coitus with an infected female. Colonization of
the entire reproductive tract with T. fetus occurs within one to two
weeks. The infection may interfere with the fertilization and devel-
opment of the resultant embryo at 50 to 70 days of gestation.
Pyometra and abortion are the first physical signs of T. fetus infec-
tion in a herd, but are believed to occur in less than 5% of infected
cows. The disease may go undetected in a herd because of its insid-
ious nature. This infection is diagnosed classically through a wet
mount or culture followed by light microscopy, neither of which
differentiates T. fetus from other trichomonads [71]. PCR assays
have recently been developed to definitively diagnose the proto-
zoan. All recently conducted PCR assays have provided new and
useful tools in the correct identification of bulls infected with T.
fetus and of bulls that may have non-pathogenic trichomonads [71,
72]. The control of bovine trichomonosis involves diagnostic test-
ing of bulls to determine their negative status prior to the breeding
season and as part of AI health programs.
Neospora caninum
Neospora caninum is a protozoan parasite that causes abortion
and serious impacts the economic performance on the dairy and
beef industries worldwide [22, 62]. Although the clinical signs of
the disease in both dogs and cattle have been recognized for over
20 years, its treatment and control options are still limited [62, 74].
The life cycle of Neospora caninum infections start with a defini-
tive host (i.e., a dog) and an intermediate host (i.e., a cattle).
Oocyts excreted from dogs may infect cattle and lead to either
infection with no discernible clinical symptoms or induce abortions
two to three weeks later, usually at about five to seven months of
gestation [73, 74]. Preventing the infection via the fecal-oral route
might not be the critical step in the prevention of Neospora abor-
tions, because oral ingestion of oocysts of N. caninum does not
reliably lead to infection and/or abortion in cattle [73]. In utero
infection of fetus from infected dams that have not aborted may be
highly efficient and a major cause of infections in the field [73, 74].
In neosprosis of cattle, both humoral and cell-mediated immune
responses have been observed in experimentally and naturally
infected animals [73, 74]. Although natural infection or vaccina-
tion produces strong antibody responses, further experiments are
required to confirm protection from abortion or vertical transmis-
sion with the antibody. To primary cattle producers, principally
three control options appear available for dealing with N. caninum
in cattle: treatment with a parasiticide that is effective against N.
caninum, a test-and-cull approach where infected animals are iden-
tified and eliminated from the herd, and vaccination [73]. Several
vaccines against N. caninum have been developed, and some of
them are commercially available [73, 74]. The vaccines consist of
inactivated vaccines, live vaccines, subunit (recombinant) vac-
cines, and vector vaccines [73].
Appropriate treatments of infected cattle followed by preventive
majors help control the disease. Effective vaccines against these
protozoans have been developed and applied in several countries.
 INFECTIOUS CAUSES OF REPRODUCTIVE DISORDERS IN CATTLES
Chlamydia spp. 10.
Since specific strains of Chlamydia pisttaci have been reported 11.
as causative agents of abortion among pregnant domestic cows in
late gestation and have also been isolated from bulls with the semi-
nal vesiculitis syndrome, several disease syndromes, including 12.
abortion and other urogenital tract infection, pneumonia, conjunc-
tivitis, enteritis, polyarthiritis, encephalomyelitis, and mastitis, as 13.
well as subclinical infections have been reported around the world
[75, 76]. Three chlamydial species that cause these conditions are
known to infect cattle: Chlamydophila (Cp.) abortus, Cp. pecorum, 14.
and the “Chlamydia-like” Waddlia chondrophila. Chlamydial 15.
abortion among cattle due to Cp. abortus occurs during the sixth to
16.
eighth months of gestation, particularly among heifers in their first
pregnancy [75]. The greatest threat is to pregnant women and
results in spontaneous abortion because of the zoonotic potential of 17.
Cp. abortus [75–77]. Recently, the involvement of Parachlamydia
spp. with bovine abortion was reported [77, 78]. 18.
Also, the association of fungus-like Aspergillus fumigatus with
bovine abortion has been reported in several regions. The fungus 19.
induces lesions on the placenta and the fetus. The abortion is
sporadic.
In addition to the aforementioned agents, several infectious 20.
agents that have not been recognized as causative agents of abor-
tion may be involved in the occurrence of reproductive disorders in
cattle. To prevent economic and/or social losses due to these disor- 21.
ders, different programs such as tests, slaughters, and vaccination
are being practiced in different country and/or regions. In conclu-
sion, the involvement of several factors, including infectious 22.
agents, indicates that both preventive and therapeutic measures are
important in minimizing the impact of reproductive disorders in
cattle. Effective preventive and therapeutic strategies must be 23.
developed and implemented, which may vary from region to region
depending on the causative agents.
24.
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