Fetal and maternal immunologic manifestations of intrauterine adenovirus SV-20 infection

Rhesus monkey fetuses of either immune or nonimmune dams were inoculated in utero with Adenovirus SV-20 (AdSV-20), a virus capable of inducing fetal pneumonia, and studied immunologically at various intervals. AdSV-20 infection at 90–100 days gestational age resulted in absolute lymphopenia in a few fetuses, reduced numbers of peripheral blood lymphocytes (PBL) which formed rosettes with sheep erythrocytes (ERL) and reduced complement-receptor lymphocytes (CRL) in a majority, while Fc fragment-receptor lymphocytes (FcRL) were occasionally increased. There was a tendency for depression of ERL and CRL early in infection of 120–130-day fetuses, followed by stimulation of these populations and FcRL in later phases. Maternal immunity did not protect against these effects of AdSV-20 infection in fetuses. Immune and nonimmune dams were spared adverse clinical effects and had no changes in lymphoid cell populations following inoculation of their fetuses.Despite precocious production of circulating IgM, fetuses of nonimmune dams had little or no demonstrable anti-AdSV-20 serum neutralizing (SN) antibody, indicating that the ability to develop an effective immune response was suppressed or had not been acquired at the gestational ages studied. Nonimmune dams displayed little evidence of seroconversion following inoculation of their fetuses with AdSV-20, except in those dams whose fetuses died in utero, whereby there was a significant antibody response. SN antibody titers of immune dams were not boostered substantially subsequent to inoculation of their fetuses, and fetal SN titers were lower than maternal titers, suggesting absence of an active fetal antibody response in this group also. Direct inoculation of AdSV-20 into 90–130-day rhesus monkey fetuses provided a model system for immunologic study of fetal infection, probably involving complex fetal-maternal interactions, in a situation where the infected, viable fetus and its dam appeared to be microbiologically isolated from one another.     

Abortion and death in goats inoculated with Sarcocystis sporocysts from coyote feces

Ten 75- to 105-day-pregnant does each were inoculated orally within 1 million (2 does), 10,000 (4 does), or 1,000 (4 does) sporocysts of Sarcocystis from coyote feces. Two does not inoculated with sporocysts served as controls. The 2 does inoculated with 1 million sporocysts died from acute sarcocystosis 21 and 22 days after inoculation (DAI), and each had 2 dead fetuses. The 4 does inoculated with 10,000 sporocysts were ill 19 to 33 DAI but survived; 1 aborted at 33 DAI, 1 had a live kid that died within 2 hours of birth 31 DAI, 1 aborted 2 dead fetuses 23 DAI, and 1 had a normal kid 56 DAI. The 4 does inoculated with 1,000 sporocysts and the 2 control does remained clinically normal and had normal kids. Does and their offspring were killed within 24 hours of parturition, and their tissues were examined histologically and microbiologically. Meronts of Sarcocystis were found in the maternal placenta of does inoculated with 1 million sporocysts. Sarcocystis was not found in the placenta, fetuses, or tissues of kids from does inoculated with 10,000 or 1,000 sporocysts, or from control does. Other abortifacient agents were not found in the placenta, fetuses, or kids from any does.     

In utero infection of swine fetuses with infectious bovine rhinotracheitis virus (bovine herpesvirus-1)

The pathogenesis of infectious bovine rhinotracheitis (IBR) virus (bovine herpesvirus-1) was studied in porcine fetuses after in utero inoculation. Laparotomies were performed on 8 seronegative pregnant sows at 34 to 86 days of gestation, and all fetuses in 1 uterine horn of each sow were exposed to IBR virus via inoculation into the amniotic sacs. Fetuses in the other horn served as controls. Clinical signs of infection were not observed in the sows, except for 2 sows that aborted at postinoculation days (PID) 11 and 15. Fetuses of the remaining 6 sows were collected at slaughter on PID 15 to 28. Fetuses were examined for gross abnormalities, presence of IBR virus in tissues, and the formation of neutralizing antibodies to IBR virus. Of 33 inoculated fetuses from 6 sows, 10 were mummified, 11 were hemorrhagic and/or edematous, and 12 were alive. Necrotic lesions were observed on the skin and in the liver of dead and live fetuses. Virus was recovered from 29 of 33 inoculated fetuses. Infectious bovine rhinotracheitis virus was isolated from fetal skin, liver, lungs, kidney, spleen, stomach contents, brain, amniotic fluid, and placenta. Virus was isolated from 4 of 11 fetuses recovered from 1 aborting sow. Antibodies to IBR virus were not detected in sera from the sows. However, antibodies were detected in 6 of 15 fetuses inoculated at 63 to 86 days of gestation and collected at slaughter at 86 to 112 days of gestation. The youngest fetus with detectable IBR antibody was estimated to be 74 days of gestation by measuring crown-rump length of the fetus.     

Attempts to establish congenital bluetongue virus infections in calves.

Three bovine fetuses were inoculated in utero with approximately 10(3) plaque forming units of type 11 bluetongue virus. The gestational ages of the fetuses at the time of inoculation were 106, 113 and 122 days. They were spontaneously aborted 104, 65 and 109 days later, respectively, and the first and third of these fetuses were recovered. There was no grossly normal cerebral tissue, the meninges formed fluid filled sacs, and the cerebellums were reduced in size. Bluetongue virus was not isolated from the fetuses but the older one had neutralizing antibody. The three dams developed neutralizing antibody to bluetongue virus. The present work supports the observation by others that early fetal infections with bluetongue virus normally result in severe central nervous system damage and not in clinically normal, persistently infected calves.     

Lesions in transplacentally induced toxoplasmosis in goats

Fifteen pregnant does were inoculated orally with 10 to 1,000 oocysts of the GT-1 strain of Toxoplasma gondii. Two does were euthanatized on postinoculation days (PID) 10 and 14, and the remaining 13 does aborted or delivered kids transplacentally infected with T gondii on PID 9 to 65. Tissues of all 34 fetuses or kids from the inoculated does were examined for gross and microscopic lesions. Placental necrosis and encephalomyelitis were the most prominent lesions. Toxoplasma gondii invaded the fetal placenta between PID 9 and 11 and invaded fetal tissues 2 to 3 days later. Necrosis attributed to toxoplasmosis, confined to cotyledons, was found in all placentas examined on PID 18. Encephalomyelitis was found in most fetuses or kids between PID 30 and 65. Lesions in fetal tissues were sparse. Because T gondii is rarely found in lesions, histologic examination of the fetal tissues is not likely to help in diagnosing every case of transplacentally induced toxoplasmosis in goats.     

Transplacental infection in mice inoculated with Getah virus

Transplacental transmission was demonstrated in pregnant mice subcutaneously inoculated with Getah virus. Viremia was shown in the infected dams, and high-titered virus was detected in the placenta and later in the fetus, suggesting virus invasion of the fetus through hematogenous infection of the placenta. High-titered virus was shown in the fetal brain and muscle and in the brain of the young dying soon after birth. Intrauterine infection resulted in a reduction of the litter size, number of young born alive and survival rate to 1 week of age. These results were further corroborated by necropsy performed several days after virus inoculation. The stage of gestation at the time of virus inoculation greatly influenced these results. Dams inoculated at 12 days of gestation delivered all dead babies, whereas virus inoculation at 5 days of gestation had no effect on the number of young born alive. The dams inoculated at 8 days of gestation had reduced litter sizes and those inoculated at 16 days of gestation produced slightly fewer live babies. Gestational stage at the time of virus inoculation also influenced viral growth in fetuses and placentas. The infection rate was low in dams inoculated at 5 days of gestation, high in dams inoculated at 8 or 16 days of gestation and 100% in dams inoculated at 12 days of gestation. High-titered virus was shown in placentas and fetuses of the dams inoculated at 8, 12 or 16 days of gestation. These results suggest that Getah virus may readily cross the placental barrier through hematogenous infection of the placenta in mice.     

Pathogenesis of porcine reproductive and respiratory syndrome virus infection in mid-gestation sows and fetuses.

Two experiments were undertaken to evaluate whether porcine reproductive and respiratory syndrome (PRRS) virus was able to cross the placenta and infect midgestation fetuses following intranasal inoculation of sows and whether PRRS virus directly infected fetuses following in utero inoculation. In experiment 1, eight sows between 45 and 50 days of gestation were intranasally inoculated with PRRS virus (ATCC VR-2332), and four control sows were inoculated with uninfected cell culture lysate. Virus inoculated sows were viremic on postinoculation (PI) days 1, 3, 5, 7 and 9, shed virus in their feces and nasal secretions, and became leukopenic. Sixty-nine of 71 fetuses from principal sows euthanized on PI day 7, 14 or 21 were alive at necropsy and no virus was isolated from any of the fetuses. Two principal sows that farrowed 65 and 67 days PI delivered 25 live piglets and three stillborn fetuses. The PRRS virus was isolated from two live piglets in one litter. In experiment 2, laparotomies were performed on five sows between 40 and 45 days of gestation and fetuses were inoculated in utero with either PRRS virus alone, PRRS virus plus a swine serum containing PRRS antibodies, or uninfected cell culture lysate. Three sows were euthanized on PI day 4 and two sows on PI day 11. Viral replication occurred in fetuses inoculated with virus alone and was enhanced in fetuses inoculated with virus plus antibody. No virus was isolated from control fetuses.(ABSTRACT TRUNCATED AT 250 WORDS)     

An experimental study on the pathogenicity of the caprine herpesvirus type 1 (CHV-1).

The pathogenicity of caprine herpesvirus type-1 (CHV-1) in goat kids and lambs was studied. Two experiments were carried out. In the first, two Saanen goat kids and four lambs of a local breed were infected intravenously with the Swiss strain E/CH of the virus. Clinical reaction was severe in the kids, but it was very mild in the lambs. Virus was excreted from the kids in higher titres and longer than in lambs. Virus was also isolated from tissue specimens but only from a kid that died on post inoculation day 4 (PID 4). The gross- and histopathological lesions were more severe in kids. In the second experiment only lambs were used. They were dexamethasone treated and then virus inoculated. A very mild infection developed. The lambs did not shed the virus, neither the virus was isolated from their tissues collected at necropsy. Nevertheless histopathological lesions were seen. In both experiments the animals seroconverted on PID 10.     

Experimental placental transfer of foot-and-mouth disease virus in mice.

An attenuated type O foot-and-mouth disease (FMD) virus which was virulent for infant, but not for pregnant, mice proved to be superior to a virulent type C FMD virus in the development of a model system for the study of placental transfer of FMD in mice. When mice were inoculated at day 8 or 12 of gestation with type O FMD virus, the virus was detectable in the maternal pancreas for 3 days and in the placenta for 6 days. Viral levels in the fetus and the amniotic fluid were inconsistent and were apparently due to a spillover from the placental infection. The elimination of the virus from the placenta coincided with the expected production of maternal 7S antibody. Mice inoculated from days 0 to 12 of gestation did not have a significant increase in dead young by day 18 (the day of necropsy). Similarly inoculated mice, when permitted to go to term, produced and raised normal-size litters. Inoculation on day 15 of gestation resulted in an increased number of deaths due to morbidity of the dams. It was concluded that the placenta serves as an active site of infection for FMD virus in pregnant mice, but the fetus is relatively resistant to infection.     

Demonstration of infectious bovine rhinotracheitis virus antigen in paraffin sections

Nine pregnant heifers were inoculated intravenously with infectious bovine rhinotracheitis virus (IBRV) in the sixth month of pregnancy. Tissues were collected from the fetus of a heifer killed 13 days postinoculation (PI), from fetuses of 6 heifers that aborted 16-27 days PI, and from mummified fetuses of 2 heifers that aborted 53 and 85 days PI, respectively. Control tissues were obtained from the fetus of a non-inoculated heifer that was killed in the seventh month of gestation. Tissues were fixed in 10% formalin, embedded in paraffin, and examined for viral antigen by immunohistochemistry, using biotinylated second antibody and alkaline phosphatase-labeled avidin-biotin complex. Antigen was detected in at least 1 tissue from the fetus of each inoculated heifer. Positive tissues included lung, liver, spleen, kidney, adrenal, and placenta. In several fetuses, antigen was identified in tissues from which virus was not isolated in cell culture. This appeared to occur when tissues had only a few small foci of infection or when tissues were severely autolyzed. The observation of viral antigen in tissues from mummified fetuses indicates that this technique may be useful in diagnostic laboratories to detect IBRV infection in tissues that are not suitable for virus isolation or for examination by the cryostat tissue section-fluorescent antibody technique.     

猪细小病毒N株的生物学和免疫学特性研究

猪细小病毒N株是从广西初产母猪所产死胎脏器分离的自然弱毒株。用这个毒株接种PPV HI抗体阴性的四月龄小猪和怀孕14～23天的后备母猪进行安全性试验,结果无任何异常临床症状、病毒血症和同居感染,母猪分娩正常,初生仔猪在吃初乳前HI抗体阴性。该毒株免疫的小猪、后备母猪和怀孕母猪,完全能抵抗猪细小病毒强毒攻击,攻毒后49天剖杀母猪,结果胎儿正常,胎儿心血HI抗体阴性,取胎儿脏器未分离出病毒,分娩母猪产仔正常,仔猪吃初乳前HI抗体阴性。而对照猪攻毒后产生病毒血症,产下不同组合异常仔,并从死胎儿脏器分离出病毒,健活仔猪吃初乳前能测出HI抗体。从而证明用N株作为弱毒苗能防止由猪细小病毒引起的繁殖障碍性疾病。     

Pathogenicity of a skin isolate of porcine parvovirus in swine fetuses

The pathogenic properties of a skin isolate of porcine parvovirus (PPV), designated Kresse isolate, were compared with NADL-8 isolate, a prototype isolate of PPV, by in utero inoculation of mid-term and late-term gestation swine fetuses. Fetuses from pregnant sows of mid-gestation were inoculated with either NADL-8 or Kresse virus. Both isolates were highly pathogenic to mid-gestation fetuses. In contrast, dramatic differences in pathogenicity between these 2 isolates were observed in fetuses inoculated late in gestation. Such fetuses from each of 4 sows were inoculated with NADL-8 or Kresse virus isolate and sacrificed at 10, 18, 21, or 23 days postinoculation (PI). NADL-8-inoculated fetuses were grossly normal. The pathogenic effects of Kresse isolate were evident by gross pathology in fetuses collected at 18, 21, and 23 days PI, but not at 10 days PI. Hemagglutination (HA) and fluorescent antibody (FA) methods were used to identify virus in various tissues of late-gestation fetuses collected at 10 and 21 days PI. At 10 days PI, HA antigens were detected only in livers of NADL-8-inoculated fetuses, but in all tissues examined of Kresse-inoculated fetuses, including the brain. PPV specific fluorescence was demonstrated in tissues of fetuses inoculated with NADL-8 and Kresse virus. The major difference was that virus antigen was found in the brains of fetuses inoculated with Kresse virus, but not in NADL-8 infected fetuses. At 21 days PI, HA antigen was not detected in any of the tissues of fetuses inoculated with NADL-8 virus, with PPV specific fluorescence by FA being found only in the kidney. However, fetuses inoculated with Kresse virus displayed HA antigen in liver and PPV-specific fluorescence in all tissues tested including the brain. Both isolates induced similar antibody responses, 1:128 to 1:256 at 10 days and 1:512 to 1:1024 at 21 days PI. In addition, immunoglobulin G (IgG) deposits were demonstrated in kidneys and skin of fetuses inoculated with Kresse virus and IgM in brain, but not in tissues from fetuses inoculated with NADL-8 virus.     

Effects on ovine fetuses of exposure to ovine progressive pneumonia virus

Fetuses of 20 pregnant ewes at 4 gestational periods (45, 55, 85, and 100 days) were inoculated with ovine progressive pneumonia virus. Fourteen of 16 fetuses exposed to virus before gestational day 80 were either resorbed or expelled, whereas 10 of 15 fetuses exposed to virus after day 80 were normal at birth. Three of the 9 expelled fetuses and 1 of 2 newborn lambs had accumulations of lymphoid cells in the lungs. Virus was readily isolated from the tissues of expelled fetuses and newborn lambs. Lambs did not have precipitating antibody to the virus at birth, but 3 to 5 lambs had specific antiviral antibody at 18 months of age.     

Congenital malformations in sheep resulting from in utero inoculation of Cache Valley virus

Serologic evidence indicated that an episode of congenital abnormalities in sheep was caused by Cache Valley virus (CVV), a bunyavirus indigenous to the United States. To determine the teratogenic potential of CVV in sheep, fetuses were infected in utero between 27 and 54 days of gestation with an isolate (CK-102) obtained in 1987 from a sentinel sheep in San Angelo, Texas. The dams of these fetuses were euthanatized between 28 and 75 days after inoculation, and the fetuses were examined for malformations. Twenty-eight of 34 fetuses had congenital abnormalities, including arthrogryposis, hydranencephaly, mummification, reabsorption, and oligohydroamnion. Virus was isolated from the allantoic fluid of 11 of 17 fetuses euthanatized at less than 70 days of gestation. The virus-positive fetuses, which were all negative for CVV-neutralizing antibody, had lesions ranging from none to severe arthrogryposis and hydranencephaly. Virus was not recovered from the allantoic fluid of fetuses after 76 days' gestation when CVV-specific antibody could be detected in 5 of 8 fetuses examined. The 2 fetuses infected on days 50 and 54 of gestation appeared normal and 1 had antibody to CVV.     

Effects of a bovine herpesvirus-1 isolate on reproductive function in heifers: classification as a type-2 (infectious pustular vulvovaginitis) virus by restriction endonuclease analysis of viral DNA

A bovine herpesvirus-1 (BHV-1) isolate (FI) from an aborted fetus was used to infect 9 heifers at various stages of gestation. Two heifers were inoculated IV on postbreeding day (PBD) 1, 7, or 14, and 3 heifers were inoculated in the sixth month of pregnancy. Plasma progesterone assays were used to monitor corpus luteum function in heifers inoculated during early pregnancy. Low progesterone values and infertility were seen in the 2 heifers inoculated on PBD 1. Luteal function remained normal in heifers inoculated on PBD 7 or 14. These 4 heifers inoculated on PBD 7 or 14 carried their fetuses to term, and their calves were free of BHV-1 infection at birth. Three heifers inoculated during the sixth month of pregnancy also carried their fetuses to term. Two calves were born alive, and BHV-1 was not isolated from nasal swab samples of either calf; the third calf was stillborn. Virus was not isolated from the stillborn calf's tissues, but BHV-1 was isolated from the placenta. Lesions were not detected in several tissues examined by light microscopy, and BHV-1 antigen was not detected by immunohistochemical examination of paraffin sections. Restriction endonuclease analysis of viral DNA was used to compare the FI virus to other BHV-1 isolates (Colorado-1, Iowa, and K22). On the basis of restriction endonuclease analysis, the FI isolate should be classified as a type-2 (infectious pustular vulvovaginitis) virus, specifically subtype a.     

A successful attempt to raise goat kids free of infection with caprine arthritis encephalitis virus in an endemically infected goat herd

Goat kids from a herd endemically infected with caprine arthritis encephalitis (CAE) virus were raised according to 3 methods. One group of ten goat kids was removed from infected does at birth before suckling or licking by the doe could occur (snatch birth technique). Kids were fed on goat colostrum, which had been heated to 57 degrees C for ten minutes and then held in a thermos flask for one hour. Subsequently the kids were fed reconstituted spray dried cows' milk powder. They were raised apart from infected goats with separation maintained by a wire fence. Contact occurred across-the-fence. Passively acquired serum antibody to CAE virus was detected in some kids at two to three months of age. Nine of the ten goats were negative for serum antibody to CAE virus when tested at 5-6, 9 and 12 months of age. One goat was positive at three and nine months of age but was negative when tested at 12 months of age. A second group of four kids was removed at birth and fed heat-treated goat colostrum, followed by milk from CAE virus-infected does. All four kids became infected with CAE virus; they developed serum antibody to CAE virus between 5-6 and 9 months of age. A third group of two kids was not removed from their infected dams. Both kids were infected at 5-6 and 9 months of age.     

Influence of maternal antibody on avian leukosis virus infection in White Leghorn chickens harboring endogenous virus-21 (EV21).

Slow-feathering (SF) white leghorn dams harboring the endogenous viral gene ev21, which encodes for complete endogenous virus-21 (EV21), and rapid-feathering (RF) dams lacking EV21 were immunized with a live field strain of avian leukosis virus (ALV) subgroup A. One group of SF dams and one group of RF dams were not immunized and were maintained to produce chicks lacking maternal ALV antibody. When the SF dams were crossed with line 15B1 males, the resulting male progeny were SF, EV21-positive, and the females were RF, lacking EV21 or congenitally infected with EV21. EV21-positive and -negative progeny of immunized and unimmunized SF and RF dams were exposed to ALV at hatching. Viremia, antibody development, cloacal shedding, and tumors in chickens lacking EV21 were compared with those in chickens with EV21. Congenital transmission of EV21 from SF dams to RF female chicks was significantly higher in immunized dams than in unimmunized dams. Maternal ALV antibody delayed infection with ALV and reduced viremia and cloacal shedding of virus in progeny. The effect of maternal antibody on ALV infection was much more pronounced in progeny lacking EV21 than in progeny harboring EV21. The data suggest that the development of ALV infection and tumors may be influenced by status of infection with EV21 and by the immune status of dams.     

Bovine Fetal Inoculations with Calf Rotavirus

The serological and histopathological responses of bovine fetuses to in utero inoculation with virulent and attenuated strains of the calf rotavirus (reovirus-like agent of neonatal calf diarrhea) are described. Thirteen bovine fetuses, 63 to 190 days of gestation, were inoculated in utero with attenuated (three fetuses) or field strain virus (nine fetuses) or both (one fetus).

Serum-neutralizing antibody titers ranging from 1:16 to > 1:256 were detected in six of eight fetuses tested, demonstrating the ability of the bovine fetus to respond immunologically to this agent. The youngest fetus in the series was inoculated at 63 days of gestation and developed a titer of 128 in 64 days. This represents the earliest stage of gestation at which a bovine fetus has been inoculated with a bovine virus and found to produce antibody to it. Serum neutralizing titers in six of the eight dams tested increased significantly following the inoculations of their fetuses in utero.

Histological changes associated with viral replication and antigenic stimulation of the lymphoreticular system were observed. Pneumonic lesions consisting of both local and diffuse lymphoreticular proliferation were present in five of the nine fetuses that were alive at slaughter. Gliosis and perivascular cuffing were noted in the brains of two of these fetuses and meningitis was seen in one. No evidence of teratogenic change was found.

     

Serological characterization of viruses isolated from experimental mucosal disease

Four cattle persistently infected with non-cytopathogenic bovine viral diarrhoea-mucosal disease (BVD-MD) virus and 3 normal controls were challenge exposed to cytopathogenic BVD-MD viruses that are antigenically different from the persistent viruses. Two of the persistently infected cattle developed mucosal disease and became moribund on postinoculation days (PID) 28 and 14, respectively; one developed severe and chronic diarrhoea and became moribund on PID 32; and the other remained healthy until the end of the experiment (PID 150). All control cattle showed transient fever, but no diarrhoea, and recovered from infection. Cytopathogenic viruses were isolated from blood of all cattle early in infection (PID 5-10) and from carcasses at necropsy. The former viruses were antigenically identical with the challenge viruses. On the other hand, the antigenicity of the cytopathogenic viruses isolated from carcasses at necropsy were different from that of the challenge viruses but similar to that of the non-cytopathogenic persistent viruses. Three of 4 persistently infected cattle, but not the calf which became moribund on PID 14, produced serum neutralizing (SN) antibodies to the challenge viruses, but not to the persistent viruses and the cytopathogenic viruses isolated from carcasses at necropsy. Control cattle produced SN antibodies to both the challenge and persistent viruses.     

Experimental respiratory syncytial virus pneumonia in young calves: microbiologic and immunofluorescent findings

Young calves were inoculated with respiratory syncytial virus (RSV) intranasally or by a combined intranasal and intratracheal route and were killed between postinoculation (initial) days (PID) 1 and 14. Viral antigens were detected by immunofluorescence in nasopharyngeal cells from calves killed between PID 2 and 10. Evidence of infection of the trachea and lungs with RSV was obtained by immunofluorescence and virus isolation in calves inoculated by the combined route, but not in calves inoculated intranasally. Within the lungs, RSV antigens were observed in epithelial cells of bronchioli and alveoli. The only virus detected in inoculated calves was RSV. With the exception of 1 calf, bacteria or mycoplasmas were not isolated from the lower respiratory tracts of inoculated calves. Antibody to RSV was not detected in calves killed up to PID 5, but 4 of 5 colostrum-deprived calves killed between PID 10 and 13 had antibodies to RSV. Preexisting, maternally derived antibody to RSV did not protect the calves from infection. Seemingly, the clinical signs of pneumonia and pathologic lesions observed in inoculated calves were caused by RSV infection.