A MUCOSAL DISEASE VIRUS AS A CAUSE OF ABORTION, HAIRY BIRTH COAT AND UNTHRIFTINESS IN SHEEP

A condition in Australian sheep resembling border disease was transmitted by the inoculation of pregnant ewes with material from affected lambs. This material contained mucosal disease virus (MDV). Twenty-two lambs comprising 6 from uninoculated control ewes, together with 11 with hairy coats and 5 with normal coats from inoculated ewes, were observed from 7 to 182 days after birth. Nine of the lambs from inoculated ewes died during the experiment from a variety of causes. Glial cell abnormalities were observed in control and affected lambs, but only 4 of the 11 hairy lambs were judged to have abnormal glial cells. There were no consistent histopathological findings indicative of MDV infection. MDV was recovered from tissues of all 11 hairy lambs, but not from any of the lambs with normal coats. The hairy lambs appeared to be immunologically tolerant to the virus. Susceptible sheep in contact with the hairy lambs were infected with MDV. It is suggested that a condition in Australian lambs characterised by hairiness of the birth coat and poor viability is due to foetal infection with a mucosal disease virus.     

TRANSMISSION OF A MUCOSAL DISEASE VIRUS INFECTION BETWEEN SHEEP

The transmission of mucosal disease virus (MDV) from infected ewes and their lambs to susceptible sheep was investigated. MDV was recovered from the amniotic fluid of an infected pregnant ewe and from the blood, nasal swabs and urine of hairy lambs. MDV infection was transmitted either at lambing, from infective foetal fluids or lambs, or later as a result of contact with a surviving with a hairy lamb and either aborted or gave birth to an infected hairy lamb. Adult sheep and 12-months-old sheep were less readily infected than were newborn lambs. Pregnant ewes were infected by contact with a hairy lamb and either aborted or gave birth to an infected hairy lamb. A method to minimise spread of the infection in the field is suggested.     

Pathology in the ovine foetus caused by an ovine pestivirus

Homogenised tissues or tissue culture supernatant fluid containing a noncytopathic pestivirus obtained from a lamb with a neurologic form of border disease, were inoculated into ewes at different stages of pregnancy. Foetal death occurred in 9 ewes of those inoculated between 19 and 47 days of pregnancy while 3 ewes did not lamb. Eight of the foetuses were aborted between 77 and 132 days of pregnancy; of these 6 were autolysed or mummified and one had arthrogryposis. The one full-term dead lamb had a hairy birth coat and lissencephalic micrencephaly. Foetal death occurred in only 7 of 14 ewes inoculated between 57 and 72 days of pregnancy. Four of these ewes aborted between 77 and 108 days of pregnancy and 3 gave birth to full-term, dead, hairy lambs. The remaining 7 ewes gave birth to live hairy lambs with severe inco-ordination. All lambs carried to term and aborted foetuses or lambs that could be examined had a range of intracranial malformations including focal leucomalacia, micrencephaly, hydranencephaly, porencephaly, lissencephaly and cerebellar hypoplasia. Some lambs also had skeletal abnormalities including arthrogryposis, scoliosis and brachygnathia inferior. The pestivirus isolate used in these trials produced more severe effects on the ovine foetus than previously observed in similar inoculation trials using pestivirus isolates from border disease lambs without nervous signs.     

A MUCOSAL DISEASE VIRUS INFECTION OF THE PREGNANT EWE AS A CAUSE OF A BORDER DISEASE-LIKE CONDITION

An experiment was designed to investigate whether a condition in Australian sheep with clinical and pathological similarites to Border Disease was caused by the infection of the pregnant ewe with a Mucosal Disease virus (MDV). Forty ewes, at 58 to 63 days after mating, were inoculated with material from lambs in which all, some or none of the tissues examined contained MDV. The clinical condition was observed only in lambs born to ewes inoculated with MDV-positive material and then only to ewes in the group which had serological evidence of MDV infection. It is concluded that the Border Disease-like condition in Australian sheep is caused by the infection of the pregnant ewe with a Mucosal Disease virus.     

The effects of bovine viral diarrhoea-mucosal disease (BVD) virus on the ovine foetus

Bovine viral diarrhoea-mucosal disease (BVD) virus has been incriminated as a cause of abortion, hairy birth coat and unthriftiness in sheep. Intravenous inoculation of 40 ewes 34 to 45 days pregnant with the V/TOB strain of virus produced death in two of four foetuses 9 days after inoculation and in all but one of 31 foetuses between 11 and 56 days. The highest levels of virus in placentomes and foetal tissues occurred between 9 and 15 days after inoculation and in foetal fluids between 11 and 18 days. Virus was not detected in any foetus later than 21 days after inoculation. Groups of 10 ewes infected between 59 and 62 days (Group B) and 70 and 76 days (Group C) of gestation had 73% and 62%, respectively, of abortions or perinatal foetal deaths. Birth weights of lambs born to infected ewes in groups B and C were significantly lower than those born to uninfected control ewes. Virus was recovered consistently from the cotyledons of the foetal membranes of live lambs, and irregularly from the tissues of full term foetuses that were dead at birth but on no occasion from mummified foetuses. There were no specific gross or microscopic lesions in tissues selected from aborted foetuses and the results highlight the difficulties associated with the diagnosis of BVD abortions and perinatal death of foetuses under field conditions.     

Effect of Rift Valley fever virus on pregnant and non-pregnant local Barki ewes after artificial infection

Studies were conducted into effects of Rift Valley fever virus on oestrous and vaginal cytology in non-pregnant ewes. 6 pregnant and 8 non-pregnant local Barki ewes were subcutaneously inoculated with different doses of virus isolated from Sharkia Province. Observed were irregularities in the oestrous cycle (20-28 days inoculation) and anoestrum. The anoestrum smears included large non-cornified epithelial cells with large well stains centrally located nuclei together with a large number of bacteria. Abortion and retention of placenta occurred in pregnant ewes with prolonged puerperal heat up to 70 days. Inoculation of virus was followed by thermal response. Re-isolation of virus in BHK cells was possible from aborted foetuses and placenta.     

Clinical variations of border disease in sheep according to the source of the inoculum

Non-cytopathogenic pestivirus obtained from lambs with border disease, with or without nervous signs, was inoculated into pregnant ewes at 57 to 65 days of gestation. Live lambs born to inoculated ewes were clinically identical to the lambs from which virus was obtained, ie, either a hairy birth coat with central nervous system disturbance or a hairy birth coat without central nervous system disturbance.     

Pathogenicity and immunogenicity of a mutagen-attenuated Rift Valley fever virus immunogen in pregnant ewes

A mutagenized clone of Rift Valley fever virus (RVFV; MV P12) used in inoculation of 3 pregnant ewes was immunogenic, nonpathogenic, and nonabortogenic. In contrast, inoculation of a matched group of 3 pregnant ewes with parent RVFV induced clinical disease and abortions. Ewes given MV P12 delivered healthy lambs that had RVFV antibody titers of less than 1:10 at birth, increasing to greater than or equal to 1:80 after ingestion of colostrum. Ewes inoculated with parent RVFV developed marked viremia, followed by RVFV antibody titers greater than or equal to 1:1,280; ewes inoculated with MV P12 developed low viremia titers and RVFV antibody titers of 1:80 to 1:320. Postpartum challenge exposure of the previously MV P12-inoculated ewes with virulent Zagazig human 501 strain RVFV indicated that the ewes were protected from clinical disease. The RVFV-susceptible female Culex pipiens that fed on the MV P12-inoculated ewes failed to transmit RVFV to hamsters; mosquitoes that fed on the parent RVFV-inoculated ewes became infected and transmitted RVFV to hamsters.     

A survey of hairy shaker disease (border disease, hypomyelinogenesis congenita) in sheep

A survey by mail in 1979 of 995 sheep farmers in Victoria enquiring as to the occurrence of hairy shaker disease (HSD) in lambs on their farms resulted in a 48% response. A total of 17 farms reported the presence of HSD. One hundred and eleven ewes produced lambs with HSD representing a small proportion only of the ewes (0.8%) on these farms and an even smaller percentage of the total ewes (0.03%) in the survey. Abortions were not prevalent on the majority of farms reporting HSD. There was no evidence that HSD was more common in any particular area in Victoria. As was anticipated mucosal disease virus/bovine virus diarrhoea (MDV) antibody was often present in serums from affected sheep. It was concluded that at present, the hairy shaker form of border disease did not appear to be an economically important problem in Victoria.     

Abortion in sheep caused by a nonclassified, anaerobic, flagellated bacterium

Twenty-eight pregnant ewes were inoculated IV with approximately 6 X 10(8) nonclassified, anaerobic, flagellated bacteria (NAFB) that had been isolated from an aborted lamb. Abortion occurred in 3 of the ewes and 1 ewe gave birth to a weak lamb. The remaining 24 ewes and 3 other ewes inoculated orally with NAFB did not develop clinical signs of illness. Suppuration and vasculitis were seen in the placentas of the 3 aborted lambs, 1 of which had necropurulent hepatitis indistinguishable from that usually attributed to Campylobacter fetus infection. The NAFB was isolated from fetal placenta, abomasal content, or internal organs of 2 aborted lambs and the weak lamb. A morphologically similar organism was seen in the abomasal content of the other aborted lamb, but the organism did not grow on bacteriologic culture medium. Therefore, in susceptible pregnant ewes, NAFB can cause fetal placentitis and hepatitis and subsequent birth of weak lambs or abortion.     

Experimental Yersinia enterocolitica placentitis in sheep.

A strain of Yersinia enterocolitica of O serogroup 6,30 isolated from the liver of an aborted ovine fetus was inoculated intravenously into a group of pregnant ewes at about 90 days gestation and produced placentitis with abortion or delivery of infected lambs about 50 days later. Y. enterocolitica of the same serogroup was recovered from the necrotic placental cotyledons and most other fetal tissues and could be isolated from vaginal discharges of the ewes for a least 2 weeks after abortion. Histological changes were consistent with an acute bacterial necrotizing placentitis and systemic infection of the fetus. Subsequent pregnancies in the ewes proceeded to term without evidence of infection.     

The production and survival of lambs persistently infected with border disease virus.

From 1985 to 1989 lambs persistently infected with border disease virus (BDV) were produced for comparative immunological studies by infecting 57 susceptible pregnant ewes between 50 and 60 days' gestation with Moredun or Oban strains of BDV. Ewes were infected either by injection with virus grown in cell culture or by housing with lambs excreting BDV. There was no significant difference in the outcomes of these different methods of infection. There was a significant difference in the number of viable lambs born to ewes receiving the two viruses. Of 41 ewes infected with Moredun virus 21 produced 32 live lambs of which 17 were reared to 1 month old (53% viability). Of 16 ewes receiving Oban virus 10 gave birth to 17 live lambs of which 15 were reared to 1 month old (88% viability). All the lambs born to ewes infected with Moredun BDV had varying signs of tremor and increased hairiness ("hairy-shakers") while those born to ewes infected with the Oban virus had no obvious clinical signs. Survival of the lambs was poor. Up until February 1991, 14 Moredun and 10 Oban sheep between the ages of 4 months and 5.5 yr had died from a variety of causes. The two commonest causes were a chronic wasting syndrome and a mucosal disease-like syndrome which was associated with the recovery of cytopathic BDV. Mating of unrelated persistently infected sheep was largely unproductive although 2 lambs were reared.     

Horizontal transmission of Aujeszky's disease virus from sheep to pigs

Eight 2-month-old merino lambs were inoculated intranasally with different (10(2.0)-10(5.0)TCID50) amounts of Aujeszky's disease virus (ADV). Electron microscopic studies indicated that ADV replicated in extra-neural sites, in the epithelial cells of the mucosa of the upper and lower respiratory tract. Although the virus was excreted continuously in nasal discharges, horizontal transmission to contact lambs failed. The surviving exposed and contact lambs had no demonstrable antibodies against ADV and they were susceptible when challenged by ADV. However, the virus was transmitted to susceptible pigs in contact with the exposed lambs. One of the five contact pigs showed characteristic clinical signs of Aujeszky's disease, developed a nonsuppurative meningoencephalomyelitis and ADV was recovered from the brain, nasal discharge and other organs. Restriction enzyme analysis of DNA from this virus confirmed the sheep origin of the isolate. The other 4 pigs seroconverted. ADV infection in sheep is therefore a possible source of infection for pigs, but the lack of horizontal transmission in sheep was confirmed.     

The effect of experimental infection with Corynebacterium pseudotuberculosis on reproduction in adult ewes

Fifteen ewes were inoculated subcutaneously with 2 x 10(6) cells of Corynebacterium pseudotuberculosis seven days before mating, during the embryonic stage of gestation and during the fetal stage of gestation. The clinical signs, antibody response and the consequences of the infection on reproduction were studied. None of the ewes showed any change in general condition during pregnancy. The effects of experimental infection during gestation were evident after incubation periods that ranged from 25 to 140 days after inoculation and were dependent on the time of the inoculation in relation to the period of gestation. Ewes inoculated before gestation can resolve infection. Ewes inoculated during the embryonic stage were severely affected and some aborted. In others lambs were stillborn or, if born alive, infected. Ewes inoculated during the fetal stage of gestation did not show reproductive disorders although some remained chronically infected.     

Observations on transplacental infection with bluetongue virus in sheep

Twenty-four ewes were inoculated with 1 of 2 strains of bluetongue virus type 4 at 40, 60, or 80 days of gestation. Two ewes aborted, 2 ewes died, and 1 was killed during the experiment, but their fetuses were recovered. At term, 2 mummified fetuses, 4 dead lambs, and 17 clinically healthy lambs were produced by 12 sheep, and the remaining 7 sheep were barren. Porencephaly and cerebellar dysgenesis were found in term lambs born to sheep inoculated at 40 and 60 days of gestation. Radiographic examination of 12 fetuses showed developmental ages far less than their chronologic age; 8 fetuses had skeletal growth-retardation lines, which were also observed in the dead lambs. A systemic lymphoreticular hyperplasia was observed in the dead lambs and in all lambs at 12 weeks of age; in 4 of the latter, granulomatous reactions were present in the liver and kidney. Lungs of the full-term lambs were reduced in weight and showed poor alveolar development and mononuclear cell infiltration, which persisted in the 12-week-old lambs. It was concluded that bluetongue virus is capable of causing not only gross abnormalities of the CNS, but also generalized growth retardation and fetal lymphoreticular hyperplasia.     

Bluetongue virus serotype 20 infection in pregnant Merino sheep

Eleven maiden Merino ewes, free of antibody to bluetongue virus serotype 20 (BTV-20) in agar gel immunodiffusion and serum neutralisation tests, were mated once with a ram. Ten ewes were inoculated with BTV-20 35 to 42 days after service, and one ewe was left as an uninoculated control. One of the inoculated ewes and the control ewe remained uninfected throughout the experiment. Eight of the remaining 9 ewes showed clinical signs ranging from mild to moderate, and the other showed no clinical signs of infection. BTV-20 viraemia was detected in ewes between days 3 and 11 post inoculation, and the serum antibody response was followed. The control ewe and 5 of the 9 infected ewes were pregnant when examined 90 to 97 days after service. Each of these animals produced a normal lamb. There was no evidence of abortion in the remaining 5 ewes, and no transplacental transfer of virus was detected in the lambs of the 5 infected ewes. At necropsy, 46 days after the birth of the last lamb, no gross or microscopic lesions were observed in either the ewes or lambs.     

Experimental infection of pregnant ewes with bovine viral diarrhea virus type-2 (BVDV-2): effects on the pregnancy and fetus

The reproduction effects of bovine viral diarrhea virus type-2 (BVDV-2) infection were investigated in ewes inoculated with a non-cytopathic BVDV-2 isolate at three stages of gestation. Virus inoculation was followed by a transient viremia, accompanied by a transient and mild hyperthermia and nasal discharge in a few animals. Some ewes were sacrificed at different time-points after virus inoculation to study the kinetics of fetal infection. Infectivity and viral antigens were detected in placentomes from day 7 to 36 post-inoculation (pi) and in fetal fluids and tissues between days 10 and 28 pi. Cardiac petechial hemorrhages and hemoperitoneum accompanied by a severe fibrinous ulcerative placentitis were observed in fetuses examined at days 21, 28 and 36 pi. Inoculation of ewes at days 55-60 of gestation resulted in a prolonged virus replication in placentomes and fetal tissues; ewes that were allowed to proceed with pregnancy had 77% of abortions or fetal and perinatal deaths. Seven stillbirths, unviable and viable lambs born to these ewes were virus-positive at birth. Infectious virus was repeatedly isolated from leukocytes of two lambs up to 2 and 6 months of age, indicating they were persistently infected. Ewes inoculated at days 65-70 of gestation had 66.6% of fetal and perinatal losses. Three viable lambs born to these ewes were healthy, BVDV antibody-positive and virus-negative. A transient viral replication in placentomes and in a few fetal tissues, followed by the rise of fetal neutralizing antibodies and virus clearance was the result of inoculating ewes at days 120-125 of gestation. Lambs born to these ewes were healthy, antibody-positive and virus-negative. These results demonstrate that the biology of BVDV-2 infection in pregnant sheep is essentially similar to that of BVDV-1 in pregnant cattle and sheep. These features make this species an attractive animal model for studying the pathogenesis of congenital BVDV-2 infection.     

Border disease in sheep caused by transmission of virus from cattle persistently infected with bovine virus diarrhoea virus.

Two outbreaks of border disease occurred on farms with sheep flocks and breeding cattle. The infection of the pregnant sheep was probably caused by transmission of virus from calves persistently infected with non-cytopathic bovine virus diarrhoea virus (BVDV) which were kept in close confinement with the ewes during mid-pregnancy. Border disease was also induced experimentally in eight lambs by exposing their dams at 38 to 78 days of gestation to a heifer persistently infected with BVDV. Both the natural and the experimental infections were characterised by typical signs such as 'hairy-shaker' lambs and high lamb mortality. The diagnosis was confirmed by virus isolations from live-born lambs, seroconversion and pathology. The study supports the assertion that cattle persistently infected with BVDV and in close contact with pregnant sheep, are an important source of strains of virus capable of causing border disease.     

Pathology of experimental Histophilus ovis infection in sheep. II. Pregnant ewes

Intravenous inoculation of Histophilus ovis to pregnant ewes at mid or late gestation resulted in acute bacterial endometritis, placentitis and foetal death. There were no lesions in the foetuses but H. ovis was recovered from the uteri of the ewes and the placentas. Oral or intravaginal administration had no effect.     

Experimental infection of pregnant ewes with enteric and abortion-source Chlamydophila abortus

Two groups of pregnant ewes were experimentally infected oronasally in midpregnancy. A faecal and an abortion-source isolate of Chlamydophila abortus were used. They were derived from a healthy ewe from a flock with no history of abortion, and from an aborted foetus in a farm with enzootic abortion. As assessed by modified Ziehl-Neelsen (MZN) staining, egg culture, antigen ELISA, the Clearview test and immunohistochemistry, inoculation resulted in placental and/or foetal infection in all ewes. Histopathology revealed placentitis in two and four ewes inoculated with the enteric or abortion-source isolate, respectively, in addition, these samples were immunohistochemically positive for chlamydial antigen. All six ewes infected with the enteric isolate and five of seven ewes infected with the abortion-source isolate showed evidence for a serological response by an indirect ELISA or CFT. Neither chlamydiae nor lesions were detected in the placentae and lambs of the uninfected control ewes, which remained seronegative. Our results suggest that enteric C. abortus can be associated with placental and foetal lesions in sheep.