Transplacental transmission of field and rescued strains of BTV-2 and BTV-8 in experimentally infected sheep

Transplacental transmission of bluetongue virus has been shown previously for the North European strain of serotype 8 (BTV-8) and for tissue culture or chicken egg-adapted vaccine strains but not for field strains of other serotypes. In this study, pregnant ewes (6 per group) were inoculated with either field or rescued strains of BTV-2 and BTV-8 in order to determine the ability of these viruses to cross the placental barrier. The field BTV-2 and BTV-8 strains was passaged once in Culicoides KC cells and once in mammalian cells. All virus inoculated sheep became infected and seroconverted against the different BTV strains used in this study. BTV RNA was detectable in the blood of all but two ewes for over 28 days but infectious virus could only be detected in the blood for a much shorter period. Interestingly, transplacental transmission of BTV-2 (both field and rescued strains) was demonstrated at high efficiency (6 out of 13 lambs born to BTV-2 infected ewes) while only 1 lamb of 12 born to BTV-8 infected ewes showed evidence of in utero infection. In addition, evidence for horizontal transmission of BTV-2 between ewes was observed. As expected, the parental BTV-2 and BTV-8 viruses and the viruses rescued by reverse genetics showed very similar properties to each other. This study showed, for the first time, that transplacental transmission of BTV-2, which had been minimally passaged in cell culture, can occur; hence such transmission might be more frequent than previously thought.     

Intrauterine transmission of ovine progressive pneumonia virus

Ovine fetuses, newborn lambs, and ovine colostrum were examined for ovine progressive pneumonia virus. The lambs and colostrum were also examined for specific antibody. Virus was isolated from 1 fetus, from 2 newborn lambs, and from most samples of colostrum. The fetus was about 100 days old and was carried by a seronegative ewe in contact with seropositive sheep. Both newborn lambs were carried by seropositive ewes. One lamb was dead at birth; the other lamb was normal and had not nursed. Antibody specific for the virus was present in the colostrum of 12 of 14 seropositive ewes and in the serum of 8 of 11 lambs that had nursed seropositive ewes, but not in the serum of lambs that had not nursed.     

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.     

Transplacental transmission of Bluetongue virus serotype 8 in ewes in early and mid gestation

The ability of Bluetongue virus serotype 8 (BTV-8) originating from the 2006 European outbreak to cross the ovine placenta during early and mid gestation was investigated in two separate experiments. In the first experiment, 16 ewes were infected with BTV-8 at 70-75 days gestation. The foetuses were collected at 18-19 days after infection (dpi). BTV-8 could be isolated from at least two organs of 19 out of 40 lambs and from 11 out of 16 infected ewes. In the second experiment, 20 BTV-8 infected ewes in early gestation (day 40-45) were euthanized at 10 days (10 ewes) or 30 days (10 ewes) after infection. The presence of BTV could be demonstrated in two foetuses from two ewes at 10 dpi and in 4 foetuses from four ewes at 30 dpi. The main pathological findings in the foetuses in mid gestation were meningo-encephalitis and vacuolation of the cerebrum. In the foetuses early at gestation, haemorrhages in various foetal tissues and necrosis and haemorrhages in the placentomes were found. These experiments demonstrate for the first time the presence of infectious BTV in lamb foetuses at different stages of gestation, combined with a difference in transmission rate depending on the gestation stage. The high transmission rate found at mid term gestation (69%) makes our model very suitable for further research into the mechanisms of transplacental transmission and for research into the reduction of this route of transmission through vaccination.     

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.     

Successful fostering of orphan lambs by nonpregnant ewes induced to lactate using exogenous hormone treatment

Six nonpregnant ewes were induced to lactate by an exogenous hormone treatment lasting 6 weeks. At the end of the treatment one alien lamb was adopted by each ewe. Each ewe was initially indifferent to the alien lamb and required only mild restraint to allow the lambs to suck successfully. Maternal interest subsequently increased and strong ewe-lamb bonding was evident by 80 minutes (n=l), 10 hours (n=4) or 24 hours (n=l) in different cases. The lambs required supplementary milk for 4–6 days while milk production by the ewes increased to adequate levels. Thereafter they were reared entirely by the ewes. Another four nonpregnant ewes were induced to lactate by a similar hormone treatment and were then hand-milked thrice daily for 2-3 weeks. One alien lamb was then introduced to each ewe. All four ewes were aggressive towards the lambs which were removed after 10–30 minutes. The ewes were then injected subcu- taneously with 5 mg of oestradiol-17β and were placed in fostering stocks. After a further 3–5 hours the lambs were re-introduced. Strong ewe-lamb bonding occurred during the subsequent 10 hours in all cases. All 10 lambs were reared successfully by these ewes until at least 3 months of age, when observations stopped. It was concluded firstly that nonpregnant ewes induced to lactate artificially will adopt and rear orphan lambs successfully, provided that additional milk is given to meet shortfalls during the first 4–6 days; and secondly that a 5 mg injection of oestradiol-17β may facilitate fostering of lambs by ewes in established lactation whether the lactation is induced artificially or not.     

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 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.     

Border Disease Virus Transmitted to Sheep and Cattle by a Persistently Infected Ewe: Epidemiology and Control

In a Swedish sheep flock comprising 202 ewes and 13 rams, a pair of twin lambs born in the spring of 1990 demonstrated signs of border disease (BD) and were persistently infected (PI) with border disease virus (BDV). Investigation showed that BDV had been introduced in the preceding tupping period 5-6 months earlier by a bought-in ewe which, on the basis of immunoperoxidase- and polymerase chain reaction techniques, was shown to be PI with BDV. Only 7 of the ewes, all of which had been in close contact with the PI ewe, seroconverted during the subsequent gestation. Apart from the PI twin lambs the losses caused by BDV were restricted to 2 barren ewes. The twin lambs, the PI ewe and lambs from the other 4 ewes that seroconverted were removed from the flock. The flock was thereafter free from an ongoing infection with BDV as shown by the absence of seroconversion.In addition, 5 heifers in late pregnancy most probably seroconverted to bovine virus diarrhoea virus (B VDV) when kept in close contact with the same PI ewe during the winter of 1989-90. When these heifers were reintroduced to the BVDV-free dairy herd from which they originated, their serum antibody titres ranged between 1:250 and 1:1250. Neither these heifers - nor their calves–caused any spread of the infection in the herd, as indicated by the absence of seroconversion in 70 cows. The present investigation shows that in the control of both BDV in sheep and BVDV in cattle, it is important to ensure that the risk of transmission of pestivirus between the 2 species is minimized.     

The effects of vaccination of Merino ewes with an attenuated Australian bluetongue virus serotype 23 at different stages of gestation

SUMMARY A cell culture attenuated Australian bluetongue virus serotype 23 (BLU23) prototype vaccine was assessed for its effects on pregnant Merino sheep. Seventy-six ewes were vaccinated at 5 different stages of gestation, and the failure to lamb at term was as follows: 35 to 43 days of gestation, 20/36 (56%); 57 to 64 days of gestation, 3/10 (30%); 81 to 88 days of gestation, 3/10 (30%); 109 to 116 days of gestation, 0/10 (0%); 130 to 137 days of gestation, 0/10 (0%). Of 30 ewes vaccinated with a cell culture supernatant fluid control between 35 and 43 days of gestation, 6.7% (2/30) failed to lamb at term. Two ewes vaccinated with BLU23 vaccine between 35 and 43 days of gestation had lambs with hydranencephaly. All other lambs born were clinically normal. Three ewes vaccinated with BLU23 aborted. Two of these were vaccinated between 35 and 43 days of gestation, the 3rd between 81 and 88 days of gestation. Five lambs were born with BLU group antibody. Four of these were from ewes vaccinated between 35 and 43 days of gestation, and 2 of these had hydranencephaly. The fifth was from a ewe vaccinated between 57 and 64 days of gestation. The vaccine did not produce disease in adult sheep, but was a potent cause of early foetal death and to a much lesser extent foetal malformation.     

Inheritance of active and acquired immunity traits in sheep

Six hundred sixteen ewes of six strains were inoculated twice with ovalbumin in Freunds' incomplete adjuvant. To quantify the humoral immune response to the foreign antigen, blood samples were collected from all ewes 1 wk post-second injection. Blood samples were also collected between 4 and 40 h of age from their 709 lambs, to examine genetic differences in ability of lambs to acquire maternal anti-ovalbumin antibodies. Titers of anti-ovalbumin antibodies were determined using kinetic enzyme-linked immunosorbent assay (ELISA) techniques. Strain did not affect ewe immune response, but sire within strain was highly significant. In pregnant ewes, anti-ovalbumin antibody titers in 12- and 30-mo-old ewes were higher than those in 21-mo-old ewes. Number of lambs in utero did not significantly affect ewe immune response. Heritabilities of anti-ovalbumin titer from a paternal half-sib analysis were .27 +/- .17 for all ewes and .57 +/- .25 for only the pregnant ewes. The effect of strain of lamb on lamb anti-ovalbumin titer approached significance, and sire within strain was highly significant. Lamb anti-ovalbumin antibody concentration increased as time from birth to blood sampling increased to 18 h but declined thereafter. The size of the litter in which a lamb was born had a highly significant effect on the lamb's acquired immunity, with titer decreasing as litter size increased. The heritability estimate for lamb anti-ovalbumin antibody concentration from a paternal half-sib analysis was .38 +/- .11; it was .28 +/- .15 from the sire variance component of a full-sib analysis. When lamb titer was considered a maternal trait (lambs nested within their maternal grandsires within strains), the maternal grandsire variance component was negative. The average anti-ovalbumin antibody concentration of lambs that died between blood sample collection and 120 d of age was less than the average antibody concentration of lambs that survived (P less than .01).     

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.     

Experimental infection of pregnant ewes with Listeria monocytogenes

Eight 18-month-old ewes were infected orally with Listeria monocytogenes between 77 and 91 days of pregnancy. Only one ewe aborted, 10 days after the first infecting dose, at 94 days of gestation; L monocytogenes was isolated from several sites in both its aborted fetuses. Two days after the first infecting dose all the ewes exhibited mild illness and pyrexia lasting for two to three days but the ewe which aborted was seriously ill until nine to 10 days after aborting. Agglutination tests carried out on 2-mercaptoethanol reduced sera revealed a strong immunological response in all the infected ewes but in the ewe which aborted this response was delayed. Four uninfected ewes which were kept as controls remained healthy throughout the experiment and showed no evidence of 2-mercaptoethanol resistant antibodies to L monocytogenes. Growth retardation lines, occurring at the time of and after experimental infection, were found in the bones of 14 of 17 newborn lambs in both the infected and control groups; in the aborted lambs these lines occurred before the infection.     

Successful fostering of orphan lambs by non-pregnant ewes induced to lactate using exogenous hormone treatment

Six nonpregnant ewes were induced to lactate by an exogenous hormone treatment lasting 6 weeks. At the end of the treatment one alien lamb was adopted by each ewe. Each ewe was initially indifferent to the alien lamb and required only mild restraint to allow the lambs to suck successfully. Maternal interest subsequently increased and strong ewe-lamb bonding was evident by 80 minutes (n=1), 10 hours (n=4) or 24 hours (n=1) in different cases. The lambs required supplementary milk for 4-6 days while milk production by the ewes increased to adequate levels. Thereafter they were reared entirely by the ewes. Another four nonpregnant ewes were induced to lactate by a similar hormone treatment and were then hand-milked thrice daily for 23 weeks. One alien lamb was then removed after 10-30 minutes. The ewes were then injected subcutaneously with 5 mg of oestradiol-17beta and were placed in fostering stocks. After a further 3-5 hours the lambs were re-introduced. Strong ewe-lamb bonding occurred during the subsequent 10 hours in all cases. All 10 lambs were reared successfully by these ewes until at least 3 months of age, when observations stopped. It was concluded firstly that nonpregnant ewes induced to lactate artificially will adopt and rear orphan lambs successfully, provided that additional milk is given to meet shortfalls during the first 4-6 days; and secondly that a 5 mg injection of oestradiol-17beta may facilitate fostering of lambs by ewes in established lactation whether the lactation is induced artificially or not.     

Evaluation of ewe and lamb immune response when ewes were supplemented with vitamin E

Fifty-two Targhee twin-bearing ewes were used in a factorial arrangement of treatments to investigate the role of supplemental vitamin E (vit E); 0 (NE) vs 400 IU of vit E x ewe x (-1)d(-1) (E) and parainfluenza type 3 (PI3) vaccination; none (NP) vs PI3 vaccination (P) in immune function. Parainfluenza type 3 vaccination was used to evoke an immune response. Ewes receiving PI3 were vaccinated at 49 and 21 d before the expected lambing date. Ewes receiving vit E were orally dosed daily, 32 to 0 d before lambing. Blood was collected from ewes at the time of the initial PI3 vaccination and 4 h postpartum. Blood was collected from lambs (n = 104) at 3 d postpartum. Ewe and lamb sera were analyzed for anti-PI3 antibody titers, immunoglobulin G (IgG) titers, and vit E concentrations. Colostrum was collected 4 h postpartum and analyzed for IgG. The model for ewe and lamb analysis included the main effects of vit E and PI3, sex (lambs model only), and their interactions. No interactions were detected (P > 0.20) for any ewe or lamb variables. Serum anti-PI3 titers were greater (P < 0.01) in P ewes and their lambs than NP ewes and their lambs. Serum vit E concentrations were greater (P < 0.01) in E ewes and their lambs than NE ewes and their lambs. Colostral IgG titers and serum anti-PI3 titers did not differ (P > 0.20) between E and NE ewes. Serum IgG titers in E ewes and their lambs did not differ (P > 0.15) from IgG titers in NE ewes and their lambs. Lamb anti-PI3 titers did not differ (P = 0.76) between lambs reared by E and NE ewes. These results indicate that, although supplemental vit E to the ewe increased lamb serum vit E concentration, it had no effect on measures used in this study to assess humoral immunity in the ewe or passive immunity to the lamb.     

Experimental intrauterine inoculation of pregnant ewes with ureaplasmas

The pregnancies of 13 ewes which were inoculated intrauterine with one of two strains of ovine ureaplasmas resulted in 9 normal and 3 abnormal births, and one ewe was found to be no longer pregnant on postmortem examination. Vaginal ureaplasma infection was detected in the majority of ewes only after lambing. Of the 12 ewes examined at postmortem, ureaplasmas were isolated from the uterus of 5 out of the 6 necropsied up to 21 days post-partum. The vulvar/preputial areas of the majority of lambs that survived were infected with ureaplasmas for the duration of the experiment, but infections of the nasal cavity and eye areas, detected at birth in 4 lambs, were resolved within 8 days post-partum.The only pathological effects detected that could possibly be attributed to ureaplasma infection were a placentitis in an ewe that aborted, and the resorption of the foetus in another.     

Effects of vitamin E and selenium injections on reproduction and preweaning lamb survival in ewes consuming diets marginally deficient in selenium

Medium wool ewes were injected with vitamin E and(or) Se over a 2-yr period to evaluate the influence of these treatments on reproduction. Ewes were divided randomly into four groups, consisting of a control, plus groups receiving monthly sc injections of either 272 iu vitamin E, 4 mg Se or 272 IU vitamin E plus 4 mg Se during pregnancy. Selenium administration increased (P less than .05) ewe blood Se concentrations, but had no effect (P greater than .10) on fertility (number of ewes lambing of ewes bred), prolificacy (number of lambs born/ewe lambing) or lamb sex ratio. Preweaning survival of lambs was increased (P less than .05) by ewe treatments with either Se or vitamin E and thus, treated ewes weaned approximately 20% more lambs/ewe mated than did control ewes.     

Effects of sarcoptic mange on the reproductive performance of ewes and transmission of Sarcoptes scabiei to newborn lambs

In each of two experiments, the reproductive performance of ewes naturally infested with sarcoptic mange and of uninfested ewes was assessed in in-season natural breeding conditions (experiment I) or in out-of-season controlled breeding conditions (experiment II). A statistically significant difference was observed in the total lambs born per ewe (infested ewes, 1.2; controls, 1.6), in the liveborn lambs per ewe (infested ewes, 1.2; controls, 1.5) and in the lamb body weight per ewe (infested ewes, 3.33kg; controls, 4.53kg) under natural breeding conditions, as well as in the lamb body weight per ewe (infested ewes, 3.37kg; controls, 4.14kg) among ewes under controlled breeding conditions. No adverse effects were observed in the other reproductive indices, cycling rate, mating rate, return-to-oestrus rate, abortion rate, lambing rate and stillbirth rate. Sarcoptic mange was observed in the offspring of infested ewes after 30 days and in the offspring of uninfested controls 60 days after their birth.     

The effects of maedi-visna virus infection on productivity in ewes

A total of five data sets obtained from four separate sheep flocks were used to evaluate the effects of subclinical maedi-visna virus infection (as determined by serological procedures) on productivity in ewes. In general, infection with maedi-visna virus had a detrimental effect on productivity. Infection resulted in a reduction in conception rates, with the odds of an infected ewe conceiving being only approximately 0.67 times those of a non-infected ewe. For ewes which did conceive, there was no detrimental effect of infection on the number of lambs born. The birth weights of lambs born to middle-aged (3–4 years) infected ewes were 3–6% lower than the weights of lambs born to non-infected ewes of the same age. There was very little apparent effect of infection in younger (1–2 years) and older (5 + years) ewes: some factors which may have biased these results are discussed. Lamb weights at 25–50 days of age appeared to be reduced by maedi-visna infection in the ewe but the results were not statistically significant.