Dual infection of fetal and neonatal small ruminants with border disease virus and peste des petits ruminants virus (PPRV): neuronal tropism of PPRV as a novel finding.

Dual infection of 26 fetal and neonatal small ruminants with border disease virus (BDV) and peste des petits ruminants virus (PPRV) is reported. The animals included five aborted lamb fetuses, 19 neonatal lambs and two neonatal kids from flocks in regions of the Black Sea and the Aegean region. BDV and PPRV antigens were detected immunohistochemically in the brain, oral mucosa, intestine and lung of infected animals. Reverse transcriptase-polymerase chain reaction was used to demonstrate PPRV and BDV in samples of the spleen, lymph node, lung and brain from infected animals. On the basis of observations made, it is concluded that brain damage following intrauterine infection with BDV facilitates the passage of PPRV to the brain and results in infection of neuronal and glial cells by PPRV.

Experimental pestivirus infections in pregnant goats.

Fifty pregnant goats, inoculated intramuscularly at different gestational stages with a non-cytopathic ovine pestivirus or a cytopathic bovine pestivirus, all developed pestivirus-neutralizing antibodies within 5 weeks of inoculation. The incidence of reproductive failure was similar for the two agents. Parturition at term with only healthy kids occurred in 13 (26 per cent) of the goats. Viable kids were not born to any of the 17 goats inoculated at about day 40 of gestation. Three of the 17 delivered dead or weak kids, seven aborted and three of seven which were necropsied during pregnancy had markedly underdeveloped and autolysed or mummified fetuses in utero, while four were barren. When inoculated at around the 60th day of gestation, two of 18 animals gave birth to only healthy kids, 12 to dead and/or weak kids, two aborted and, at necropsy, a small, decomposed fetus was found in one goat while one other was barren. In this group, one kid was ataxic and seven others had body tremors characteristic of border disease. One of the latter kids was viable. Of 15 goats inoculated at around day 100 of gestation, 11 gave birth to healthy kids only, three to dead and/or weak kids and one aborted. In 23 progeny, histological changes in the central nervous system (CNS) consisted mainly of cerebral white matter necrosis, cerebellar dysplasia, hypercellular areas in white matter and lymphocytic perivascular cuffings. All seven weak-born kids with signs of border disease had CNS lesions, particularly cerebellar dysplasia and/or hypercellular areas. Non-cytopathic pestivirus was isolated from tissues from all eight progeny examined in the 40-day inoculation group, from tissues and/or serum from 10 of 23 progeny in the 60-day group, and from four of 24 in the 100-day group. Persistent infection was demonstrated in a healthy kid, in a viable shaker and in two other kids which appeared normal at birth. Examination of offspring before ingestion of colostrum revealed pestivirus antibodies in one kid in each of the 40- and 60-day inoculation groups and in five kids in the 100-day group.

Border disease. Virus-induced decrease in thyroid hormone levels with associated hypomyelination.

Border disease (BD) was induced in lambs by inoculation of their dams at 50 days gestation with Border disease virus (BDV) isolate #31. At birth, the clinically affected lambs had diffuse spinal cord hypomyelination, confirmed by immunocytochemical staining for myelin-associated glycoprotein and myelin basic protein. In the BD lambs, large numbers of thyroid follicular epithelial cells and scattered pituitary cells contained BDV antigen by immunofluorescence staining. Double labeling techniques demonstrated the BDV-infected pituitary cells to contain growth hormone, adrenocorticotrophic hormone, prolactin, or luteinizing hormone. Cells containing thyroid stimulating hormone were rare and were not positive for BDV antigen. Infection of the pituitaries and thyroid glands caused no detectable morphologic changes as compared to controls. The BD lambs had statistically significantly (p less than 0.05) lower mean serum concentrations of thyroxine and L-3,3',5-triiodothyronine as compared to age-matched uninfected controls. Similar significant differences in the mean plasma levels of growth hormone and thyroid stimulating hormone were not found. In addition, the BD lambs had a statistically significant (p less than 0.05) lower mean activity of the myelin-associated, thyroid hormone dependent enzyme, 2',3'-cyclic nucleotide-3'-phosphodiesterase in spinal cord tissue. Although not conclusive, these results indicate that the hypomyelination in BD may be due to depressed levels of circulating thyroid gland hormones secondary to a noninflammatory and noncytolytic infection of the thyroid gland by BDV. This is one of the first reports indicating that a virus-induced hormonal alteration may cause a congenital lesion in animals.

Experimentally induced ovine border disease: extensive hypomyelination with minimal viral antigen in neonatal spinal cord.

Border disease (BD) was experimentally induced in 9 lambs by inoculation of their dams with the BD-31 strain of border disease virus (BDV) at 50 days of gestation. These ewes developed serum-neutralizing antibody titers to BDV. The diagnosis of BD in their lambs was confirmed by hairy birthcoats, intrauterine growth retardation, tremors, and hypomyelination. Three clinically healthy age-matched control lambs, whose dams had been given an inoculum containing only BDV-free cell culture supernatant, were studied in parallel. There were significant differences in birth weights and in the lengths of the right tibiae and radii between the affected and the control lambs. There was a gradient in severity of clinical neurologic signs among the affected lambs, which directly correlated with the severity of hypomyelination in their spinal cords. However, the difference in severity of the neurologic deficits did not correlate with differences either in the precolostral serum-neutralizing antibody titers to BDV in these lambs or in the number of BDV antigen-containing cells in their spinal cords. Only approximately 0.01% to 0.3% of spinal cord cells, both in gray matter and white matter, were BDV antigen positive in the affected lambs. These results indicate that extensive infection of CNS cells with their subsequent destruction or functional alteration may not be a critical part of the pathogenesis of the hypomyelination in BD.

Viral persistence and abnormalities of the central nervous system after congenital infection of sheep with border disease virus.

Lambs congenitally infected with border disease (BD) virus and sheep exposed to BD virus as adults were studied for one year to determine the pathogenesis of congenital exposure compared with adult exposure to the virus. Persistent BD virus was isolated in tissue culture and detected by immunofluorescence of the peripheral white blood cells, urine, and cerebrospinal fluid of lambs with congenital BD up to one year of age. These animals had no detectable serum neutralizing antibody response to the virus for the same interval. BD virus antigen was also detected by immunofluorescence in many central nervous system tissues of these lambs with congenital BD. Dysmyelination and glial proliferation in the central nervous system and microencephaly were noted in the lambs with congenital BD, and these lesions appeared to remain the same over a 12-month period.