Colostral transmission of maedi visna virus: sites of viral entry in lambs born from experimentally infected ewes

Maedi visna virus (MVV) vertical transmission in sheep via infected colostrums is a very important route of infection in lambs. To verify colostral transmission and to study early viral entry in lambs, colostrum samples, and small intestine and mesenteric lymph nodes of lambs born from experimentally infected ewes were examined by histopathology, immunohistochemistry (IHC) and in situ hybridisation (ISH) studies. In particular, newborn lambs were naturally fed maternal colostrum and humanely killed at 10, 24, 48, 72, 96 h and 7 and 10 days after birth; two caesarian-derived lambs served as uninfected controls. No lesions suggestive of MVV infection were found, but marked immunoreactions for MVV capsid antigen (CA, p28) were detected in lambs fed maternal colostrum and in macrophages cultured from colostrum. IHC results in lambs suggest an initial viral absorption by intestinal epithelial cells at the tip of the villi, passage to mononuclear cells in the lamina propria and involvement of ileum Peyers' patches and mesenteric lymph nodes, with different staining patterns depending on infection times. ISH on intestinal sections of the 72 h lamb revealed the presence of proviral DNA in epithelial cells at the tip of the villi, suggesting a role for these cells in early MVV replication. The results contribute to knowledge about the pathogenesis of ovine lentivirus infection suggesting that the small intestine and mesenteric nodes are the sites of entry and propagation of MVV in lambs fed colostrums from infected ewes.     

The effect of a natural maedi-visna virus infection on the productivity of South African sheep

A cohort study was conducted in order to measure the effect of the chronic indurative lymphocytic mastitis caused by the South African strain of maedi visna virus (MVV) on the pre-weaning growth of lambs born either of naturally infected or uninfected ewes kept under similar conditions. Fifty naturally infected ewes as well as another 40 from a maedi-visna-free source to be used as control animals, were purchased and kept in separate flocks which were managed in a similar way. All the ewes were of the same breed and 3-4 years old. During the adaptation period, and through the mating, pregnancy and lactation periods they were periodically monitored for the presence of MVV serum antibodies. The lambs were weighed at birth and thereafter every 2 weeks until the age of 90 days, when they were weaned. The ewes were then slaughtered, and their udders examined histologically and the number of lymphocytic follicles were counted and assessed. Although the calculated values indicated a correlation between the number of follicles in the udder and the reduction in the growth rate of the lambs, this was not statistically significant. Similarly, despite higher counts of lymphoid follicles in the udders of sero-positive ewes as compared to those that were sero-negative and the lower ewe productivity indexes in infected ewes, no statistically significant differences were found in the indexes of ewes in different follicle categories.     

Studies in epidemiology of maedi/visna in sheep.

Lambs born to ewes from flocks with a high incidence of maedi/visna were separated from their dams at birth (group 1), or after 10 h (group 2), six weeks (group 3) or one year (group 4) and were observed for periods of up to eight years. Group 1 lambs remained free of infection while 28 per cent, 76 per cent and 81 per cent respectively of lambs in the other groups developed clinical, serological or histopathological evidence of infection during the observation period. It is therefore concluded that vertical transmission, if it occurs at all, is of little significance in the epidemiology of the disease. The number of serologically, virologically and histopathologically maedi/visna positive sheep, the time of onset of disease and the severity of lesions were related to the duration of exposure to the parent flock. In a separate trial no evidence was obtained for the transmission of maedi/visna by Muellerius capillaris larvae.     

Protection of lambs against enteric colibacillosis by vaccination of ewes

Pregnant ewes were vaccinated twice, seven weeks and three weeks before lambing, with a multivalent formalin-killed Escherichia coli vaccine containing an added K99, F41 antigen preparation. Lambs born to vaccinated and unvaccinated ewes were exposed to oral infection with E coli B44 (09:K30, K99, F41). All 10 lambs from vaccinated ewes were protected whereas all 10 control lambs developed severe diarrhoea and five died or were killed in extremis. In the following year, previously immunised ewes were given a single dose of the vaccine two weeks before lambing. Eleven of their 12 lambs were protected against a similar challenge, which caused the death of six of eight control lambs and severe diarrhoea in the two survivors. Higher levels of antibody to the K99, F41 preparation were detected by enzyme-linked immunosorbent assay in the serum and colostrum from vaccinated ewes and in the serum of their lambs when compared with similar samples from control ewes and lambs.     

Longitudinal studies of naturally acquired Brucella abortus infection in sheep.

Naturally acquired Brucella abortus infections were studied during consecutive pregnancies in eight sheep and in their lambs over a period of 40 months to evaluate epizootiologic aspects of natural infection in sheep. Brucella abortus was isolated from the ewes following 16 of 26 natural terminations of pregnancy: from 5 of 6 ewes in the first year, from six of eight ewes in the second year, from two of six ewes in the third year, and from three of six ewes in the fourth year. Vaginal swab samples and milk samples were the most consistent source of the brucella organisms. Brucella abortus was isolated from three ewes when standard tube test seroagglutination titers were less than 1:100. In contrast, results of supplemental tests (card, 2-mercaptoethanol, complement-fixation, and Rivanol) remained positive during the study. During the 40 months, B abortus was isolated from 4 of 4 aborted fetuses, 2 of 5 stillborn lambs, 10 of 37 living lambs, and as an indicator of continuing infection, from 6 of 12 lambs born during the fourth year. Although B abortus has a definite host preference for cattle, this study demonstrated that under appropriate management conditions, sheep may be naturally infected and may remain infected for more than 40 months. Epizootiologic evaluation of all factors, including husbandry practices and exposure potential, should be utilized in determining the need to test other species that may have been exposed to cattle infected with B abortus.     

Effect of passive transfer of maternal immune components on infection with ovine herpesvirus 2 in lambs

OBJECTIVE: To define the role of passively tranferred immunity in protection against early infection with ovine herpesvirus 2 (OvHV-2) in lambs. ANIMALS: 15 adult sheep and 34 lambs. PROCEDURES: 2 groups of animals were used, including 15 lambs born to OvHV-2-free ewes and 19 lambs born to OvHV-2-positive ewes. After nursing colostrum, all lambs and their dams were introduced into a flock positive for OvHV-2. Blood was obtained from the lambs every 2 weeks and examined by PCR assay and competitive inhibition ELISA. RESULTS: None of the animals had positive results by PCR analysis for samples obtained approximately 2 weeks after introduction into the flock. In the group of lambs from OvHV-2-infected ewes, 5 of 19 had positive results at 1 month of age and 17 of 19 by 5 months of age. In the group of offspring from OvHV-2-negative ewes, only 1 of 15 had positive results at 1 month of age, and the number reached 12 of 15 by 5 months of age. All lambs in both groups had positive results by 6 months. An active antibody response to the virus was detected in animals within 3 weeks after viral DNA became detectable in the blood. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis suggests that passively transferred immunity does not play an important role in the delay of infection with OvHV-2 in lambs. Age also does not seem to influence susceptibility. The rate of infection in young lambs may simply be a reflection of the intensity of viral exposure in their environment.     

Vertical transmission of Toxoplasma gondii in naturally infected ewes in the semiarid region of Brazil

To evaluate transplacental transmission of Toxoplasma gondii in naturally infected ewes, blood samples were collected from 55 pregnant ewes and their offspring, before ingestion of colostrum. From 16 offspring of positive ewes and nine offspring from negative ewes, blood samples were obtained after 48 h and 7, 14, 21, 28, 35, 42, 49 and 56 days after birth. T. gondii antibodies were detected in serum samples using the indirect fluorescence antibody test (IFAT ≥ 64). Four of the 30 positive ewes (13.3 %) had offspring positive for T. gondii before ingesting colostrum (vertical transmission). The colostrum antibody titers decreased every week, and only 20 % (2/10) of the lambs in continued to present detectable antibody titers until day 56 after birth. Therefore, vertical transmission of T. gondii in lambs was indication of occur and is an important route for transferring and maintaining the agent in sheep herds in the Brazilian semiarid region.     

Relative contribution of colostrum from Maedi-Visna virus (MVV) infected ewes to MVV-seroprevalence in lambs

Maedi-visna virus (MVV) seroprevalence associated with consumption of colostrum from seropositive ewes was investigated in 276 housed lambs from birth to 300 days-old. At birth, lambs were allocated to five experimental groups according to the maternal MVV-serological status, source and mode of feeding colostrum (bovine or ovine and bottle fed or suckled from the dam) and type of horizontal MVV-exposure (raised with the dam or separately with other lambs). The risk of being seropositive at 300 days-old was associated with feeding ovine colostrum from seropositive ewes and increased with intake of bottle-fed ovine colostrum and was higher in lambs separated from their dams and raised with other experimental lambs compared to lambs raised with their dams. Approximately 75-87% of ELISA-positive results in lambs that had ovine colostrum was attributable to colostrum itself. However, approximately only 16% of naturally raised and 29-61% of bottle-fed ovine colostrum lambs were ELISA-positive as a result feeding ovine colostrum. These results confirm that ovine colostrum from seropositive ewes can be a major source of MVV but its overall contribution to seroprevalence in natural conditions is relatively low, and shows that horizontal MVV transmission can be an important source of infection in new-born lambs.     

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.     

Seroreactivity of Peruvian sheep and goats to small ruminant lentivirus-ovine progressive pneumonia virus

Sera from 3,369 sheep and 1,394 goats in Peru were examined by agar-gel immunodiffusion for antibodies to ovine progressive pneumonia virus (OPPV). The point prevalence rates for antibodies to OPPV in sheep were 1.7% to 40.6% (mean, 19.02%) in the 7 flocks studied, whereas for goats, the point prevalence rates for antibodies that cross-reacted with OPPV in 12 herds were 0.0% to 45.1%. For sheep, a direct association between increasing age and increasing seroreactivity to OPPV was established, and there was evidence to indicate that lambs born to primiparous ewes and raised separated from all other sheep after they were weaned may have been less likely to become infected with OPPV than those lambs born to multiparous ewes and not separated from other sheep after they were weaned. For goats, antibodies to OPPV were detected in 7 of 12 herds studied, the highest infection rate being present within a herd in the Lima department (district).     

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.     

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.     

The effect of subclinical experimental Cowdria ruminantium infection in ewes on the growth and milk consumption of pre-weaning lambs.

An alternative control option for heartwater (Cowdria ruminantium infection) is the establishment and maintenance of endemic stability which would lessen the existing dependence on acaricides. In an endemically stable state, animals become infected by vaccination or natural challenge at an early age, following which the immunity so created is boosted by continuing tick challenge. In this study, growth rates, health and hematological parameters were monitored at regular intervals for lambs born to two matched groups of ewes until weaning at 4 mo of age. One group of ewes was infected multiple times with Cowdria ruminantium; the other group remained uninfected. The overall mean leucocyte count of lambs born to infected ewes was significantly lower than that of lambs born to uninfected ewes (P=0.04). However, there were few other significant differences in the other hematological data between the two groups. The mean birth weight of single lambs born to uninfected ewes (4.6 kg) was significantly higher than the mean birth weight of single lambs born to infected ewes (4.4 kg) (P=0.02). Trends in milk consumption and growth rates were similar for the two groups, with few significant differences detected. Likewise, there were no significant differences in the incidences of health problems or pre-weaning mortalities between the two groups of lambs. The results of this study indicate that there is no detectable effect on productivity of pre-weaning lambs when their dams are carriers of C. ruminantium--a situation likely to occur in an endemically stable state. Hence, maintenance of endemic stability would be a suitable control option for heartwater.     

Maedi-visna control in sheep. I. Artificial rearing of colostrum-deprived lambs

A field trial to study the practicability and efficacy of maedi-visna control in sheep by artificial rearing of lambs was carried out during the lambing season of 1979. Lambs were immediately separated from the dams at birth, deprived of ovine colostrum, and reared isolated from the parent flock. Bovine colostrum was given instead of maternal colostrum. Eleven farms participated in the experiment. All flocks were severely infected with maedi-visna virus: 63-100% of the ewes were seropositive as demonstrated by ELISA. Artificially reared lambs were serologically tested and positives culled at the age of 6, 12, 18, 24, 30 and 36 months. Only very few positives were found: 1/389, 1/376, 0/337, 1/223, 1/192 and 0/144, respectively. The first two sero-positive lambs occurred in one flock, and it could be ascertained that both had mistakenly been given ovine colostrum probably containing maedi-visna virus. No explanation, other than sub-optimal hygiene and isolation, could be found for the two sero-positive sheep that turned up in another flock at 24 and 30 months of age although, transplacental infection cannot be entirely excluded. It is concluded that artificial rearing of ovine colostrum-deprived lambs is an effective and practicable method for the control of maedi-visna in sheep. The method appears particularly useful when valuable genetic material has to be salvaged.     

Vitamin E status in newborn lambs with special reference to the effect of dl-alpha-tocopheryl acetate supplementation in late gestation

Pregnant ewes were supplemented with dl-alpha-tocopheryl acetate, either as a single intramuscular dose (500 mg two weeks before lambing) or perorally (150 mg daily during 3-4 weeks before lambing). Ewes without such a supplementation were controls. The vitamin E supplemented ewes had nearly twice as high vitamin E (alpha-tocopherol) concentrations as the unsupplemented control ewes at lambing both in serum and in colostrum. The vitamin E concentration in colostrum was 5-11 higher than in milk 1 week after lambing. Both supplementations somewhat increased the vitamin E serum concentration of the newborn lambs, but the increase was negligible in comparison with the effect produced by the consumption of colostrum. All lambs had very low serum concentrations at birth. The lambs from the supplemented ewes had significantly higher serum values than the control lambs 24 h after birth. The ewes had somewhat higher selenium status at birth than their offsprings when evaluated by glutathione peroxidase (GSH-Px) in the erythrocytes. It seems reasonable that nutritional muscular degeneration may arise in newborn lambs with a normal selenium status if their vitamin E status is critical, either because of an inadequate consumption of colostrum or because of a vitamin E deficient diet during pregnancy with a low vitamin concentration of colostrum as a consequence.     

Transmission and control of caprine arthritis-encephalitis virus

Caprine arthritis-encephalitis virus was isolated from goat milk and transmitted most efficiently to kids through both milk and colostrum. In addition, it appeared that transmissions through other secretions of the doe occurred, but were less important than transmission in milk and colostrum. Intrauterine infection may have occurred in 2 of 32 cesarean-derived goats, but postpartum horizontal transmission could not be ruled out. Transmission by the aerosol route was not demonstrated, and even short-term direct contact between virus-infected bucks and virus-free does during breeding did not result in transmission. Prolonged direct contact for over 12 months between weaned cesarean-derived goats and virus-infected goats was necessary before horizontal transmission could be demonstrated under nondairy conditions. However, when uninfected does were milked with infected does, a high percentage became infected in less than 10 months. Heat inactivation (56 C) reduced approximately 10(5) median tissue culture infective doses of caprine arthritis-encephalitis virus to below titratable levels, and virus was not transmitted to kids fed virus-infected colostrum that had been heated at 56 C for 1 hour. A program of eradication is discussed in which kids are removed from their dams at birth, fed safe sources of colostrum and milk, and isolated from other goats until weaning.     

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.     

Does sward height grazed by ewes in mid- to late-pregnancy affect indices of colostrum intake by twin and triplet lambs?

AIMS: To determine if sward height grazed by ewes in mid- to late-pregnancy affects subsequent indices of colostrum intake by twin and triplet lambs, and the extent of the variation in colostrum intake between twin and triplet litter-mates.

METHODS: Blood samples were collected from 75 twin and 46 triplet litters, respectively, 24–36 h after birth. Samples were not collected from lambs that died before this age. Lambs were born to ewes which had grazed one of four sward heights (2, 4, 6 and 8 cm) during mid- to late-pregnancy. Serum samples were analysed for gamma-glutamyl transferase (GGT) activities and glucose concentrations. The number of lambs present at weaning was used to determine lamb survival rate.

RESULTS: GGT activities of lambs born to ewes that grazed swards 2 cm high were lower than those born to ewes that grazed swards 4 and 6 cm high (p=0.053 and p=0.037, respectively). Glucose concentrations were higher in twin than triplet lambs (p=0.003) and in lambs born to ewes grazed on swards 6 cm high than those born to ewes grazed on swards 2 cm high (p=0.033). These differences were not evident after correction for differences in birthweight. There was significant (p<0.001) variation within litter-mates for GGT activities and glucose concentrations in both twins and triplets. The blood constituent status of lambs that died before the 24–36 h collection period is not known. However, lambs which died after this period had significantly lower GGT activities (p=0.009) and glucose (p=0.010) concentrations at the time of sampling than those which survived.

CONCLUSIONS: Farmers should ensure twin- and tripletbearing ewes in mid- to late-pregnancy are grazing at least 4 cm of herbage to ensure colostrum intake of lambs in the postpartum period is not adversely affected.     

Pilot project for eradicating maedi-visna in Walliser blacknose sheep

Maedi-Visna is a lentiviral disease of sheep with a worldwide distribution. The transmission of the virus occurs primarily via colostrum and milk from the infected ewe to its newborn lamb but also horizontally between sheep. The most obvious clinical symptoms are progressive dyspnea and emaciation. In this prospective study an eradication based on serological testing and removing of seropositive animals was performed in 24 flocks of sheep of the breed "Walliser Schwarznasenschafe" leading to a reduction of the seroprevalence from 36% to 1% within two years. The control group consisted of 21 flocks of sheep. Lambs of seropositive ewes had a 7.6 times higher risk to seroconvert within their first two years of life compared to those of seronegative ewes. The dynamics of the spread of the infection were studied in birth cohort groups. Cohort animals of seropositive ewes showed an obvious trend to seroconvert slowly. Seropositive ewes had a significantly lower reproduction rate and their lambs suffered from significantly higher death and lower growth rates, probably due to a reduced milk production, resulting in economic losses.     

Epidemiological studies on a hairy shaker virus infected flock

A serological study of hairy shaker disease (HSD) was carried out on a group of 30 Romney two-tooth ewes and their lambs over a two year period. Although ten ewes showed seroconversion to HSD virus during pregnancy, an association between infection with HSD virus and reproductive failure was not established. Passive immunity was demonstrated in nine of 21(43%) of the lambs. The duration of passive immunity was 10-20 weeks (mean 14.9 weeks) with the half life of antibody being 18.7 days (SD=3.04). The lambs were studied serologically for 12-20 months. Infection with HSD virus was demonstrated in nine of 16 lambs (56%) with most infections occurring in the summer and autumn when they were three to eight months old.