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1.
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.  相似文献   

2.
Adenovirus SV-20 (ASV-20) was inoculated subcutaneously into adult rhesus macaques (Macaca mulatta), and various immunologic parameters were studied. Similar changes were observed in macaques that had anti-ASV-20 serum-neutralizing antibodies prior to inoculation and in those lacking detectable antibodies. There were absolute decreases in numbers of peripheral blood lymphocytes (PBL), erythrocyte-rosetting lymphocytes, complement-receptor lymphocytes, and Fc-receptor lymphocytes. These changes were most significant (P less than 0.05) on postinoculation days (PID) 3 and 7. Mitogenic responsiveness to concanavalin A, phytohemagglutinin, and pokeweed mitogen in cultured PBL from immune and nonimmune macaques was depressed on PID 3, 7, and 14. Ultraviolet-inactivated ASV-20 caused moderate suppression of phytohemagglutinin-induced mitogenesis when viral particles and lectin were added simultaneously to PBL cultures. Plasma cortisol (hydrocortisone) values were not significantly altered following inoculation of ASV-20. High titers of anti-ASV-20 antibody developed by PID 7 in nonimmune macaques, and previously immune macaques showed a booster effect in the same time period. Antibody titers were still increased 120 days after inoculation. There was no clinical evidence of an adverse effect of ASV-20 infection in these macaques.  相似文献   

3.
Thirteen pregnant goats were inoculated intravenously with the ALD strain of virulent swine fever (SF) virus on Days 64-84 of gestation. Dams showed transient and mild viremia, and produced high serum neutralizing (SN) antibody after inoculation. Six inoculated dams were reared until parturition occurred and bore six apparently normal, one apparently normal but dead, one mummified and three edematous kids. Neutralizing antibody was demonstrated in the pre-colostral sera obtained from all normal kids, but no SF virus was isolated from any of them. The other seven dams were killed on post-inoculation days (PID) 5-61, and fetuses, placenta and amnion were tested for the virus and SN antibody. All fetuses of five dams examined within PID 40 were positive for SF virus, but negative for SN antibody. SF virus was also isolated from one of three fetuses examined on PID 61. Conversely, the other two fetuses examined on PID 61 were negative for SF virus, but positive for SN antibody. Furthermore, SF virus was isolated from the placenta and amnion of all the dams.  相似文献   

4.
Porcine fetuses were exposed in utero to porcine reproductive and respiratory syndrome virus (PRRSV) at stages of gestation ranging from 34 to 85 days and examined 17 to 31 days later to determine the effect of gestational age on fetal susceptibility. For each of the 8 litters tested during the study, all of the fetuses of 1 horn of the uterus were exposed to virus by intraamniotic injection; those of the other horn were exposed similarly to a sham inoculum that consisted of sterile cell culture medium. Viral infectivity titers associated with fetal tissues collected at necropsy indicated that, regardless of gestational age, the virus had replicated in fetuses exposed intraamniotically. In addition, virus had also spread and replicated in sham-inoculated littermates in 3 litters. On the basis of these findings it appears that there may be little or no temporal difference in fetal susceptibility to infection with PRRSV. If so, the lack of early fetal death as a commonly recognized feature of naturally occurring cases of PRRS may be due to a greater resistance of early gestational fetuses to the lethal effects of PRRSV, as suggested by this study, and/or a greater likelihood of transplacental infection during late gestation.  相似文献   

5.
The serological and virological results of an experimental infection of bovine fetuses with bovine viral diarrhea virus are presented. Four fetuses, 120-165 days gestational age, were inoculated in utero with a second passage virus strain. Two fetuses received a sham-inoculum. A humoral immune response in the virus-inoculated fetuses, was demonstrated three weeks later. In three fetuses only IgM and IgG1 were detectable. The serum from the fourth fetus also contained IgG2 and IgA. Bovine viral diarrhea virus-neutralizing antibodies were detected in two fetuses. These two fetuses inoculated at 135-150 days gestational age, represent the youngest reported bovids, giving a specific response in three weeks following an experimental infection with bovine viral diarrhea virus. The fetal sera did not contain heat-labile factors, which could mediate the neutralization. The virus was not reisolated from any of the fetuses, but viral antigen was nevertheless demonstrated by immunocytochemical methods in sections of several of the fetal organs, primarily lymphoid tissues.  相似文献   

6.
Immune potency test was conducted in horses by inoculating a killed vaccine for equine viral arteritis (EVA) which had been freeze-dried and contained aluminum hydroxide adjuvant. Serum neutralizing (SN) antibody to equine arteritis virus (EAV) was detected at maximal titers of 1:80 to 1:640, 1 to 2 weeks after 2-dose vaccination of 6 female horses. However, 6 pregnant mares inoculated with the vaccine which had been kept in storage for 1 year at 4°C produced much higher titers ranging from 1:320 to 1:1280. A maximal mean titer of 1:199.5 occurred in the 1st and 2nd week after 2-dose inoculation with the nonpreserved vaccine, whereas a maximal mean titer of 1:794.3 occurred in the 2nd week using the preserved vaccine. The horses showed no systemic or local adverse reactions clinically or hematologically after vaccination. Four of the 6 vaccinated pregnant mares were exposed to the Bucyrus strain of EAV but resisted challenge exposure, while 3 nonvaccinated control pregnant mares revealed acute EVA causing abortion and death. Isolation of EAV was positive from the body tissues of the aborted and dead fetuses and their dams, but was negative from the vaccinated mares. No significant rise of SN antibody titers was detected in the vaccinated mares following challenge exposure, suggesting that the vaccine can protect against EAV infection in pregnant mares and prevent abortion or death.  相似文献   

7.
The aim of this study was to characterize the pathogenesis of Neospora caninum in experimentally inoculated pregnant water buffalo (Bubalus bubalis). Twelve Mediterranean female water buffaloes ranging in age from 4 to 14 years old and seronegative to N. caninum by indirect fluorescent antibody test (IFAT) were involved. Ten females were intravenously inoculated with 10(8) tachyzoites of NC-1 strain at 70 (n=3) or 90 (n=7) days of pregnancy (dp). Two control animals were inoculated with placebo at 70 and 90 dp, respectively. Serum samples were obtained weekly following inoculation to the end of the experiment. Three animals inoculated at 70 dp were slaughtered at 28 days post inoculation (dpi), three animals inoculated at 90 dp were slaughtered at 28 dpi and the remaining four animals inoculated at 90 dp were slaughtered at 42 dpi. Fetal fluids from cavities and tissue samples were recovered for IFAT and histopathology, immunohistochemistry and PCR, respectively. Genomic DNA from fetal tissues was used for parasite DNA detection and microsatellite genotyping in order to confirm the NC-1 specific-infection. Dams developed specific antibodies one week after the inoculation and serological titers did not decrease significantly to the end of the experiment. No abortions were recorded during the experimental time; however, one fetus from a dam inoculated at 70 dp was not viable at necropsy. Specific antibodies were detected in only two fetuses from dams inoculated at 90 dp that were slaughtered at 42 dpi. No macroscopic changes in the placentas and organs of viable fetuses were observed. Nonsuppurative placentitis was a common microscopic observation in Neospora-inoculated specimens. Microscopic fetal lesions included nonsuppurative peribronchiolar interstitial pneumonia, epicarditis and myocarditis, interstitial nephritis, myositis and periportal hepatitis. Positive IHC results were obtained in two fetuses from dams inoculated at 70 dp and slaughtered at 28 dpi. N. caninum DNA was detected in placentas and fetuses from all inoculated animals. The pattern of amplified microsatellites from placental and fetal tissues resembled the NC-1 strain. Water buffaloes, like cattle, are susceptible to experimental inoculation with N. caninum at early pregnancy.  相似文献   

8.
Four-week-old pigs of the Chester White, Duroc, Hampshire, Landrace, and Yorkshire breeds (n = 518) were vaccinated with a pseudorabies modified live-virus vaccine to determine whether genetic differences existed for immune response after vaccination. All pigs and their dams (sows) were tested before vaccination to determine preimmunization antibody titers, using a microtitration serum-neutralization (SN) test. The SN test results of sows were negative, as were preimmunization tests of the pigs. At 4 weeks after pigs were vaccinated, additional blood samples were collected from the pigs, and end-point SN titers were determined, using a 2-fold dilution scheme. Small, but statistically significant, breed differences existed for antibody response, with Yorkshire and Chester White pigs having the highest response, and Duroc and Landrace pigs, the lowest. Differences among sire progeny groups were small, but there were significant differences among dams. Genetic differences as seen by differences among breeds indicates that the efficacy of vaccines may vary from breed to breed and that vaccine trials should not neglect this potential source of variation.  相似文献   

9.
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.  相似文献   

10.
Transplacental infection of hamster fetuses was produced by inoculation of pregnant hamsters with 10(6.3) plaque-forming units (PFU) of Akabane virus by either the intraperitoneal or the subcutaneous route. Virus with titers as high as 10(7.5) PFU/g of tissue was detected first in the placenta and later in the fetus. Virus could also be readily isolated from blood, lung, spleen, and liver of both pregnant and nonpregnant hamsters, but it reached higher titers and persisted longer in the placenta and fetus. Young dying at birth had Akabane virus titers as high as 10(7.3) PFU/g of brain tissue. Litter size was reduced by inoculation of the pregnant hamster at gestational day 11 or earlier, and survival of the newborn to 1 week of age was decreased by inoculation at gestational day 9 or later.  相似文献   

11.
Minute Virus of Canines (MVC) was shown to cause transplacental infections with embryo resorptions, especially in dams given parenteral inoculations prior to gestational day 30. Responses to oral-nasal inocula were inconsistent. Direct exposures of embryos or fetuses to MVC via the amniotic sac, however, resulted in their deaths after an incubation period of approximately 2 weeks. Virus was prominent in fetal lung and small intestinal villi, but it could not be demonstrated in fetal or uterine tissues greater than 2 weeks after the estimated time of embryonic/fetal death. With 1 exception, dams inoculated during the last trimester of pregnancy gave birth to normal pups; some pups had developed active immune responses. The exceptional dam whelped dead or dying pups with anasarca or myocarditis. Antibody levels to MVC were generally higher in dams with infected fetuses than in bitches with normal pups at term. Failures to isolate virus, or demonstrate viral antigens in tissues by immunostaining, other than near the time of embryo/fetal death, indicate the need for more sensitive viral detection methods.  相似文献   

12.
The Nc-Spain 7 isolate of Neospora caninum, which was newly obtained from an asymptomatic congenitally infected calf, demonstrated a similar virulence as Nc-1 strain in mouse models. The aim of this study was to characterize the pathogenesis of Nc-Spain 7 isolate in cattle after experimental infection at 65 days of gestation. For this purpose, thirteen pregnant heifers were divided into three groups as follows: group A: 7 heifers inoculated with 1×10(8) tachyzoites of Nc-Spain 7 isolate; group B: 4 heifers inoculated with 1×10(8) tachyzoites of Nc 1 strain; and group C: 2 heifers received PBS. Serum samples were collected weekly and heparinized blood samples were collected three times (0, 28 and 42 days after inoculation) by jugular venipuncture. Placenta and fetal tissue samples were collected at time of necropsy. Specific antibody response in the dams was tested by IFAT, indirect ELISA, and rNcGRA7 and rNcSAG4 based-ELISA. Specific antibody response in fetal fluids was tested by IFAT. IFN-γ production was measured after in vitro culture of PBMC and the supernatant was assessed using a commercial kit (BOVIGAM). A significant increase in N. caninum antibody responses was detected in groups A and B by IFAT and by i-ELISA from day 14 after inoculation onwards. Besides, antibody response against rNCGra7 protein was also detected in all inoculated heifers by rNcGra7-based ELISA. Four fetuses from group A and one from group B were aborted between 3 and 5 weeks after infection. In the recovered fetuses, only 3 out of 4 fetal fluids from fetuses of group A and 1 out of 3 of group B were seropositive by IFAT, but all of them were positive by PCR. Transplacental transmission could be determined in all fetuses from groups A and B by PCR and/or IHC. Heifers of group C and their fetuses remained negative by all techniques. The results of this study demonstrate that the NC-Spain 7 isolate could be transmitted transplacentally, and produced fetal death and abortion in cattle.  相似文献   

13.
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.  相似文献   

14.
From a colony of 131 owl monkeys, adenovirus was isolated 68 times from 433 oropharyngeal and rectal swab sample pairs collected over a 2-year period. A total of 63 of 68 adenovirus isolates were grouped by neutralization into 1 of 3 types designated owl monkey adenovirus types (OMAV Ty) I, II, and III. The OMAV Ty I and Ty II were identified by neutralization with antiserum to squirrel monkey adenovirus type I and adenovirus SV-11, respectively. The OMAV Ty III was partially neutralized by antiserum to OMAV Ty II. The OMAV Ty I was isolated from 8 of 30 newly imported owl monkeys within 3 weeks of arrival. A total of 13 of 26 owl monkeys had a fourfold or greater rise in serum-neutralization (SN) titer with 25 of 27 developing SN titers of greater than or equal to 1:80. A total of 40 of 87 other owl monkeys in the colony had SN titers of greater than or equal to 1:5. The OMAV Ty II was isolated from 51 (12%) of 433 swab samples pairs. Virus was isolated from 26 owl monkeys with 13 having persistent infections. Viral isolations spanned a 10- to 17-month period for five of these owl monkeys. In 114 owl monkeys, 101 (86%) had SN titers to OMAV Ty II greater than or equal to 1:20. The OMAV Ty III was isolated from three owl monkeys. Of 114 owl monkeys, 93 (82%) had SN titers to OMAV Ty III of greater than or equal to 1:20. Although the distribution of SN titers were similar for OMAV Ty II and Ty III, 42 (37%) of 114 owl monkeys had eightfold or greater differences in SN titers between OMAV Ty II and Ty III.  相似文献   

15.
Change of porcine circovirus 2 (PCV2) target cells during development from fetal to postnatal life in pigs was examined. PCV2 inoculation was performed in fetuses in utero at either 57, 75 or 92 gestational days and in piglets at 1 day of age. Twenty-one days after virus inoculation, PCV2-infected cells in the heart, lungs, liver, spleen and inguinal lymph nodes were localized and immuno-phenotyped by double-immunofluorescence labeling using different cell markers and PCV2-antibodies. During fetal life, viral antigens were detected in cardiomyocytes, hepatocytes and macrophages and infected cell numbers decreased with increasing fetal age at inoculation. The heart contained the highest number of infected cells and cardiomyocytes were the main target cell. Postnatally, macrophages were the only target cell type in different organs and infected cell numbers were similar to those of fetuses inoculated at 92 days of gestation. One piglet showed exceptionally high number of infected cells in different organs with values 13-513-fold higher compared to littermates. In this piglet, the majority of infected cells in lymphoid tissues could not be typed. This study reveals that PCV2 target cells change from cardiomyocytes, hepatocytes and macrophages during fetal life to only macrophages postnatally.  相似文献   

16.
Fetal serum from most of 994 bovine and 553 ovine aborted fetuses was tested serologically for antibodies to border disease (BD), bovine viral diarrhea (BVD), and bluetongue (BT) viruses, and to Leptospira sp., and the results were compared with the results of isolation procedures, fluorescent antibody tests (FAT), and histologic examinations of the same fetuses. Antibodies to BT virus were not found in any of the 994 bovine and 553 ovine fetuses. Antibody titers to BVD virus were present in 39 of 966 bovine fetuses tested, and BVD virus was detected in 4 of the 39. Four of 74 fetuses in which the BVD virus was detected by FAT or isolation had titers to BVD virus. Microagglutination (MAT) titers to 1 or more of 5 serovars of leptospires were present in 52 of 773 bovine fetal sera tested. Leptospires were not detected by FAT in any bovine fetuses that had leptospiral antibody titers. Leptospires were detected by FAT in 15 aborted calves, and none of these had MAT titers. Antibody titers to BD virus were present in 80 of 486 fetal lamb sera tested, and the virus was detected by FAT or isolation in 3 of the 80 fetuses. Border disease virus was detected in 14 of 486 fetal lambs tested. Twelve of the 14 were tested serologically and 3 had titers to BD virus. Leptospiral antibody titers were present in 27 of 326 ovine fetal sera tested. Leptospires were not detected in any of the 326 ovine fetuses tested by FAT.  相似文献   

17.
Antibody responses of hens and their progeny were studied in commercial broiler nuclear lines. Starting at 168 days of age, individually housed pullets from lines A and B were fed a 16% crude protein and 2752 kcal metabolizable energy/kg mash diet supplemented with either 10 or 300 IU/kg of vitamin E fed as dl-alpha-tocopherol acetate. Fifty-eight days later (226 days of age), 12 hens per line-vitamin E subclass were inoculated i.v. with 0.1 ml of a 2.5% suspension of sheep red blood cells (SRBC). Plasma antibody titers were measured 6, 20, 40, 54, 70, and 88 days after inoculation. Hens from both lines were artificially mated to males from line C, and progeny from eggs collected 9-15, 25-30, and 65-70 days after inoculation were tested for antibodies to SRBC. Hens were reinoculated i.v. with 0.1 ml of 0.25% SRBC 88 days after the first inoculation, and their antibody levels were measured 3, 6, and 20 days later. Eggs laid 10-13 days after reinoculation were incubated, and antibody titers of chicks were measured at hatch. Antibody response of hens to an initial inoculation of SRBC was line-diet-time after inoculation specific. In line A, titers were greater for hens fed the lower than the higher vitamin E diet, whereas diet had no effect on the antibody levels in line B. Line effects (A > B) were observed on days 6 and 20 after inoculation but not thereafter. After the second inoculation, dietary vitamin E level had no effect on antibody levels of hens within lines, whereas a between-line difference (A > B) was observed for the lower but not the higher level of dietary vitamin E. Although there was no difference between diets for antibody transferred to progeny by line B, there was a difference (lower > higher) for line A. After reinoculation of their dams, antibody titers of chicks from line A, but not line B, reached levels similar to those after the first inoculation. Antibody levels were higher for chicks at hatch than in 16-day embryos or 10 days posthatch. The results of this research suggest genetic variation in response to immune stimulation by dietary vitamin E.  相似文献   

18.
Nineteen pregnant (45 to 90 days of gestation) and 9 nonpregnant ewes were inoculated orally with 1,000 or 10,000 oocysts of Toxoplasma gondii. Pregnant ewes were euthanatized at days 14 (2 ewes), 21 (1 ewe), 23 (1 ewe), 28 (2 ewes), 35 to 42 (6 ewes), and 49 to 62 (6 ewes), and antibody titers in fetal and maternal sera were assayed, using the modified agglutination, latex agglutination, indirect hemagglutination, and dye tests. Although all ewes developed antibody titers of greater than or equal to 1,024 within 28 days after inoculation, fetuses were seronegative up to 28 days, using the modified agglutination test. Toxoplasma gondii antibodies were found in fetuses, using the modified agglutination and dye tests 35 days after ewes were inoculated. Latex agglutination and indirect hemagglutination tests were insensitive for detection of T gondii antibodies in ovine fetal sera. Toxoplasma gondii antibody titers in nonpregnant ewes were similar to those in pregnant ewes. Passively acquired T gondii antibodies from the colostrum decreased from 1,024 to less than 16 between 49 and 56 days of age in 1 lamb and between 62 and 106 days in its twin.  相似文献   

19.
The objective of this study was to demonstrate the efficacy of a modified-live virus (MLV) vaccine in protecting fetuses from infection with type 1 or type 2 Bovine viral diarrhea virus (BVDV) when pregnant heifers were challenged at approximately 170 d of gestation with noncytopathic field isolates. The 83 pregnant heifers had been bred naturally 4 wk after vaccination. Fetuses were collected 60 d after BVDV type 2 challenge, and newborn calves were collected before colostrum intake after BVDV type 1 challenge. Protection was determined by measuring the serum neutralizing (SN) antibody response in the fetus or calf and by virus isolation from thymus, lung, spleen, and kidney tissue samples. There was a measurable SN antibody response to BVDV in all the fetuses and calves of the control heifers, which had received a placebo vaccine. However, only 4 of 22 calves and 7 of the 28 fetuses of the MLV-vaccinated heifers demonstrated SN antibody after BVDV challenge. Type 1 BVDV was isolated from tissue samples of 5 of the 12 calves of control heifers and none of 22 calves of the MLV-vaccinated heifers challenged with type 1 BVDV. Type 2 BVDV was isolated from tissue samples of 17 of the 18 fetuses of the control heifers and 2 of the 28 fetuses of the MLV-vaccinated heifers challenged with type 2 BVDV. The results of this study demonstrate that the MLV vaccine reduces the fetal infection rate by at least 82% for BVDV type 1 and by 75% for BVDV type 2 when heifers are exposed to highly fetotrophic BVDV at 170 d of gestation.  相似文献   

20.
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.

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