Transplacental infection of porcine fetuses following experimental challenge inoculation with encephalomyocarditis virus.

Ten multiparous sows were inoculated between 46 and 50 days of gestation with a fetal swine isolate of encephalomyocarditis virus (EMCV) to investigate the ability of the virus to cause transplacental infection and fetal death. Four sows (group 1) were inoculated IM with EMCV MN-25 that had been passaged 4 times on baby hamster kidney-21 line cell monolayers. Two sows were euthanatized at postinoculation (PI) day 23, and the other 2 sows at PI day 44. An additional 6 sows (group 2) were inoculated IM with the same virus that had been passaged 5 additional times in pigs. Two sows were euthanatized at 14 days, and the remaining 4 sows at PI day 28. Clinical signs were not observed in any of the sows, whereas all sows seroconverted to EMCV. In group 1, only 2 of 50 fetuses were mummified. Virus was not recovered, although EMCV antibodies were detected in the 2 mummified fetuses. In group 2, the 2 sows that were euthanatized at PI day 14 had 26 normal fetuses and there was no evidence of fetal infection. However, in the 4 sows euthanatized at PI day 28, 20 of 48 fetuses were mummified, hemorrhagic, or edematous. Encephalomyocarditis virus was recovered from 21 of 48 fetuses. Transplacental infection and fetal deaths in pregnant sows was achieved following infection with EMCV passaged in pigs.     

Porcine parvovirus: propagation in microcarrier cell culture and immunogenic evaluation in pregnant gilts

Porcine parvovirus was propagated in PK-15 cells cultured in roller bottles or on microcarrier beads. After inactivation, the virus was used as antigen in the preparation of vaccines. The immunogenic potency and safety of the vaccines were evaluated in specific pathogen free pregnant gilts and guinea pigs. Experimental challenge tests determined the efficacy of the vaccine in preventing porcine parvovirus transplacental infection. Neither viral antigens nor specific antibodies were detected in fetuses from vaccinated gilts. In contrast, fetal death and, or, mummification occurred when unvaccinated gilts were infected. Both virus and, or, antibodies were also detected in fetuses from these unvaccinated gilts. Serum conversion after vaccination was assayed by microserum neutralisation using guinea pig erythrocytes as cell indicators and by haemagglutination inhibition tests. Viral antigens in fetal tissues were detected using ELISA, the immunobeads technique, the haemagglutination test and by virus isolation.     

An Experimental Infection With Classical Swine Fever Virus in Pregnant Sows: Transmission of the Virus,Course of the Disease,Antibody Response and Effect on Gestation

An experimental infection with classical swine fever (CSF) virus in 12 conventional gilts, housed in a sow‐box housing system, was conducted in order to evaluate horizontal transmission, clinical, virological and serological response, and the effect on gestation. Two of the 12 gilts, of which 10 were pregnant, were experimentally inoculated. They became viraemic for the first time 6 days post‐inoculation (dpi). The contact gilts became viraemic between 18 and 21 days post‐inoculation. On the basis of virological findings and the martingale estimate of R₀ (13.0) it was concluded that the two experimentally inoculated gilts infected all contact gilts, although random contacts between gilts were not possible. The presence of CSF infection could be diagnosed earlier and during a longer period when the leucocyte count or polymerase chain reaction were used in comparison with virus isolation in whole blood (P < 0.05). The observed clinical symptoms were atypical and highly variable between the gilts, which hampered clinical diagnosis. The pregnant gilts became infected between day 43 and 67 of gestation. In all cases vertical virus transmission occurred and this resulted partially in abortion and/or mummification.     

Viremia and effect of fetal infection with porcine viruses with special reference to porcine circovirus 2 infection

This publication reviews some pathogenetic features of the transplacental infection with porcine viruses in sows. Viremia either with virus freely circulating or associated to peripheral blood mononuclear cells (PBMC) is an essential part of such pathogenesis. Virus replication occurs either in fetal tissues only or both in fetal and maternal tissues and the outcome may be different.Since porcine circovirus 2 (PCV2) has been associated with reproductive failure in sows, the question was asked what type of viremia PCV2 causes and what the effect of PCV2 is on the pregnant uterus. Seronegative gilts were oronasally inoculated and plasma and PBMC were monitored for infectious virus and for quantity of viral DNA copies. Infectious virus was found in plasma only at 21 days post-inoculation (DPI). Virus associated to PBMC was detected between 14 and 49 DPI. Viral DNA was found in plasma between 14 and 49 DPI and associated to PBMC between 7 and 63 DPI (end of experiment). Direct intra-fetal inoculation at 57, 75 and 92 days of gestation and collection of fetuses 21 days later showed that the virus replicates highly in fetal tissues, particularly in the heart. Fetal death occurred in the 57 days sows while virus and antibodies were observed in the 75- and 92-day inoculated sows. Inoculation at 57 and 75 days of gestation and collection of the piglets at the end of pregnancy showed that intrauterine spread had occurred to fetuses adjacent to the inoculated ones and that fetal death occurred also in the presence of antibodies. The pregnancy was not interrupted.This study shows that PCV2 causes viremia which is largely cell-associated and that virus replication in fetuses causes fetal death with mummification. Whether such transplacental infection occurs in the immune sow population is questionable.     

Infectivity of porcine circovirus type 2 DNA in semen from experimentally-infected boars

Porcine circovirus type 2 (PCV2) is an economically important pathogen. It has been demonstrated that PCV2 DNA can be detected in boar semen by PCR; however, the biological relevance of this is unknown. The objectives of this study were to determine if semen positive for PCV2 DNA is infectious (1) in a swine bioassay, or (2) when used for artificial insemination. For the first objective, 4-week-old pigs were inoculated intraperitoneally with PCV2 DNA-negative (bioassay-control; n = 3), PCV2a DNA-positive (bioassay-PCV2a; n = 3), or PCV2b DNA-positive (bioassay-PCV2b; n = 3) raw semen, or PCV2 live virus (bioassay-positive; n = 3), respectively. Pigs inoculated with PCV2 DNA-positive semen and PCV2 live virus became viremic and developed anti-PCV2 antibodies indicating that the PCV2 DNA present in semen was infectious. For the second objective, three Landrace gilts were inseminated with PCV2 DNA-negative semen (gilts-controls) from experimentally-infected boars, and six gilts were artificially inseminated with semen positive for PCV2a DNA (gilts-PCV2a; n = 3) or PCV2b DNA (gilts-PCV2b; n = 3). Serum samples collected from the gilts in all groups remained negative for anti-PCV2 antibodies for the duration of the experiment. In addition, fetal serum samples from all 105-day-gestation fetuses were negative for anti-PCV2 antibodies or PCV2 DNA. Under the conditions of this study, PCV2 DNA-positive semen was not infectious when used to artificially inseminate gilts; however, it was demonstrated to be infectious in a swine bioassay model and therefore is a potential means of PCV2 transmission amongst swine herds.     

Classical swine fever virus strain "C" protects the offspring by oral immunisation of pregnant sows

The aim of this study was to evaluate if oral immunisation of wild sows protects the fetuses from transplacental infection. Two experiments were carried out with gilts vaccinated orally with C-strain virus approximately 5 weeks after insemination. They were challenged at mid-gestation with highly virulent classical swine fever virus (CSFV) or moderately virulent field virus. The results revealed that oral vaccination has no negative impact on the pregnancy, and all vaccinated sows developed neutralising antibodies. After infection no symptoms were detected in the six vaccinated-infected sows. Challenge virus could neither be found in blood, nasal and fecal swabs or saliva nor in organs sampled at necropsy. Likewise, all fetuses originating from vaccinated sows were virologically and serologically negative. In contrast, the controls developed a short viremia and as a result of the transplacental infection all fetuses were CSFV positive. In addition, 22 serologically positive wild sows of an endemically infected area, where oral vaccination had also been carried out, and their offspring were free from CSFV or viral RNA. Our results confirm that oral immunisation of pregnant wild sows with C-strain vaccine may protect the fetuses against CSF.     

Bovine viral diarrhea infection in pregnant swine

Twenty pregnant gilts (5 groups of 4) were infected experimentally with 1 of 4 strains of bovine viral diarrhea virus (BVDV) administered intranasally-orally. Blood specimens were taken from the gilts on postinfection day (PID) 7 and cultured for virus. Serum specimens, obtained on PID 21 and at termination of the experiment, were tested for neutralizing antibodies. At 90 to 112 days of gestation, the gilts were euthanatized and their fetuses were examined for evidence of intrauterine infection. Evidence of infection was demonstrated in all of the gilts, either by isolation of BVDV at PID 7 or subsequently by detection of neutralizing antibody titers. Intrauterine infection was confirmed in one of 20 gilts by isolation of BVDV, detection of neutralizing antibodies, and demonstration of microscopic lesions in the fetuses. The microscopic lesions were characterized as nonsuppurative meningitis and choroiditis. Clinical signs of disease were not seen in the infected fetuses. Of 8 gilts exposed to strains of BVDV pathogenic for cattle, 1 gilt developed intrauterine infection, 2 gilts were found barren, and 3 gilts had significantly fewer fetuses than copora lutea.     

An inactivated, oil-emulsion vaccine for the prevention of porcine parvovirus-induced reproductive failure

Pig fetuses inoculated at 45 days gestation with virulent porcine parvovirus (PPV) were harvested 10 days later. Virus was extracted, inactivated with binary ethylenimine and the antigen suspension emulsified with mineral oil adjuvant. One dose of this vaccine, or two doses with a 14 day interval, stimulated high and long lasting serum antibody titres in gilts. Vaccination caused no clinical reactions and lesions at injection sites were minor. Vaccination of seronegative gilts at 40 days gestation caused no adverse effects on fetuses. Six gilts which had been vaccinated five to nine weeks before mating were challenged intravenously with live, virulent PPV at 40 days gestation. At 98 days gestation 78 out of 84 (93 per cent) fetuses were alive and normal and no evidence of PPV infection was found in the six dead (mummified) fetuses. In four unvaccinated gilts similarly challenged with PPV at 40 days gestation only five out of 51 (10 per cent) fetuses survived to 98 days gestation and the virus was detected in 41 of the 46 dead (mummified) fetuses. This vaccine appears to be safe and effective for prevention of PPV-induced fetal loss in gilts.     

The effect of breed and intrauterine crowding on fetal erythropoiesis on day 35 of gestation in swine

In a previous report, it was suggested that intrauterine crowding impaired fetal erythropoiesis and that fetal erythropoiesis was accelerated in Meishan pigs during early pregnancy. Because these conclusions were based on limited numbers of observations, the present experiment was undertaken to provide a more extensive investigation of these phenomena. Intact white crossbred gilts, unilaterally hysterectomized-ovariectomized (UHO) white crossbred gilts, and intact Meishan gilts (n = 13 to 16 per group) were mated after at least one estrous cycle of normal duration (17 to 23 d). Gilts were laparotomized at d 35 of pregnancy, the uterine horns were exteriorized and opened near each fetus, and a blood sample was collected from each fetus. The uterine horn was then surgically removed, and each fetus and placenta was weighed. All fetal blood samples were measured for hematocrit, red blood cell number, and hemoglobin. Erythropoietin and the percentages of nucleated cells and reticulocytes were also measured in blood samples from the largest and smallest living fetus in each litter. Fetal hematocrits were not affected by treatment. Blood cell counts were greater (P < 0.01) in fetuses of Meishan gilts than in White crossbred intact or UHO gilts. Hemoglobin was less (P < 0.01) in fetuses of Meishan gilts than in fetuses of White crossbred intact or UHO gilts. The percentages of nucleated (immature) cells and reticulocytes were both less (P < 0.01) in fetuses of Meishan intact gilts. Erythropoietin was also lower (P < 0.01) in fetuses of Meishan gilts. As observed previously, fetal weight was correlated (r = 0.38; P < 0.01) with blood hemoglobin concentration. These results confirm that fetal erythropoiesis in Meishan gilts is accelerated compared with White crossbred gilts. These results are consistent with the hypothesis that faster blood cell development could be beneficial to fetal survival in swine.     

Effect of number of conceptuses on maternal hormone concentrations in the pig

This study examined the effect of number of conceptuses on maternal concentrations and profiles of estrogen sulfate, estrone, estradiol-17 beta, progesterone and prolactin in gilts. Estradiol-valerate injections were used to induce pseudopregnancy (O conceptuses; n = 5) and oviduct ligation or no treatment were utilized to obtain pregnant gilts with 4 to 7 (n = 4), or 8 to 11 (n = 4) conceptuses, respectively. Blood samples were collected every 10 d from d 10 through 110 of pregnancy or pseudopregnancy. At 110 d after onset of estrus, all gilts were slaughtered and numbers and(or) weights of fetuses, corpora lutea, placentae and the empty uterus were determined. Concentrations of estrogen sulfate and estrone, but not progesterone or prolactin, were associated with fetal number, total fetal weight, total placental weight or empty uterine weight. In contrast, only progesterone was highly correlated with number of corpora lutea. Results suggest that most conjugated estrogen, estrone and estradiol were of fetal-placental origin, whereas little, if any, placental production of progesterone or prolactin occurred. Increases in estrogen sulfate and estrone concentrations were observed at gestation d 30 and from d 70 to 100. The latter increase coincides with previously established increases in the rate of maternal mammary development and fetal growth.     

Effects of zearalenone (F2) on estrous activity and reproduction in gilts

The effects of zearalenone on swine reproduction were investigated in two trials involving a total of 82 gilts which were allotted into three groups at puberty, mated at second estrus and slaughtered 80 d postbreeding. A control diet without mycotoxin (group 1) or an experimental diet containing 3.61 ppm (first trial) or 4.33 ppm zearalenone (second trial) were fed at a mean daily level of 2 kg/animal. The experimental diet was fed from puberty to mating (group 2) or during pregnancy (group 3). No difference was observed between the two trials. When fed to nonpregnant gilts, zearalenone induced a pseudopregnancy state in 45% of the animals; no estrus was detected within 50 d following puberty and corpora lutea developed at puberty were maintained. The uterine horns were edematous. Reproductive performance measured at 80 d postmating (ovulation rate, weight of corpora lutea, number of normal and abnormal fetuses, embryonic mortality) were not affected by zearalenone intake. But when zearalenone was fed during pregnancy, weights of uterus, placental membranes and fetuses were significantly decreased in comparison with those of control gilts and heterogeneity of fetuses in the same litter was increased. Hematocrit and erythrocyte count were lower in fetuses from gilts ingesting zearalenone, but hematology of the dams remained unaffected. No mycotoxin residue could be detected in gilts or fetal tissues despite the great consequences observed on cyclicity of the females or on the development of embryos. This experiment showed evidence of the estrogenic properties of zearalenone in mature gilts.     

Hepatitis E Virus Antibodies in Finnish Veterinarians

We investigated hepatitis E virus (HEV) infections in Finnish veterinarians engaged in different practice specialties and evaluated the effect of different background factors on HEV exposure by examining total HEV antibodies in samples collected from the participants of the 2009 National Veterinary Congress in Helsinki, Finland. Finnish veterinarians commonly have total HEV antibodies with seroprevalence of 10.2%. Of the non‐veterinarians, 5.8% were seropositive. Increasing age was associated with HEV seropositivity, and, surprisingly, the highest HEV seroprevalence (17.8%) among veterinarians was detected among small animal practitioners. Although no positive correlation between swine contacts and HEV seropositivity was found, 22.7% of veterinarians who had had needle stick by a needle that had previously been injected into a pig versus 9.0% of those who had not were seropositive, even though the finding was statistically non‐significant (P = 0.07). Our results suggest that, although contact with swine is a known risk factor for HEV infection, the sources of HEV infections are probably numerous, including travelling abroad and possibly also other reservoirs of HEV than pigs.     

Cross-protection studies between feline infectious peritonitis and porcine transmissible gastroenteritis viruses

Cross-protection studies between the feline infectious peritonitis (FIP) and the porcine transmissible gastroenteritis (TGE) viruses were conducted in cats, pigs and pregnant gilts. Cats vaccinated with TGE virus developed neutralizing antibodies against TGE virus and low titer antibody against FIP virus detected by an indirect fluorescent antibody technique but were not protected against a virulent FIP virus challenge. Baby pigs and pregnant gilts vaccinated with FIP virus did not develop detectable antibodies to TGE virus. Nevertheless, it appeared that vaccination of swine with FIP virus conferred some immunity against TGE virus infection. Seventeen-day-old pigs vaccinated with two doses of FIP virus had a 67% survival rate following a virulent TGE virus challenge, and 75% of the 3-day-old pigs suckling either FIP or TGE-virus-vaccinated gilts survived virulent TGE virus infection in contrast to 0% survival of baby pigs suckling unvaccinated gilts.     

Association of hepatitis E virus (HEV) and postweaning multisystemic wasting syndrome (PMWS) with lesions of hepatitis in pigs

The aim of the study was to determine the presence of swine hepatitis E virus (HEV) RNA and antibodies in postweaning multisystemic wasting syndrome-affected (n=114) and non-affected (n=46) pigs and the possible association with hepatitis lesions. Forty-four pigs were RT-PCR positive (28.2%); 25 of them were PMWS cases, while 19 were non-PMWS pigs. In both groups, HEV RT-PCR results were associated with hepatitis (OR=5.61 for PMWS-affected pigs and OR=5.17 for non-PMWS affected pigs; p=0.01). No interaction was detected in a logistic regression between PMWS occurrence and HEV infection for the development of hepatitis lesions. Seropositivity to HEV was more likely to occur in pigs with hepatitis (51.9%) compared to pigs without hepatitis (36.1%; p=0.03). Significant differences in optical densities were notices comparing the lesional stage of pigs (p=0.009). While pigs with slight or moderate hepatitis were seropositive, pigs with more severe lesions were seronegative to HEV. These results indicate that swine HEV infection can be a significant contributor to the development of moderate hepatitis in pigs regardless of the PMWS status.     

Pigs orally inoculated with swine hepatitis E virus are able to infect contact sentinels

The purpose of the present study was to explore the most likely natural route of infection of swine hepatitis E virus (HEV) by oral inoculation of pigs and to investigate the potential infection by direct contact exposure. A preliminary experiment was performed to assess the infectiousness of the bile used as source of virus. Once confirmed, 16 pigs were inoculated via oral drop with an HEV positive bile suspension containing 2 × 10⁵ genome equivalents per pig. Nine animals were kept as contact sentinels and 12 more pigs were used as negative controls. A number of pigs from the three groups were euthanized at 16, 32 and 64 days post-inoculation. From the HEV inoculated group, three pigs shed virus in faeces, two had virus RNA in bile at necropsy and two seroconverted. In the contact group, two animals showed presence of HEV RNA in bile. This study demonstrates that pigs orally inoculated with a single HEV dose got infection, although few animals had evidence of infection. Moreover, the virus was successfully transmitted to direct contact exposed pigs.     

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。     

Localisation of swine hepatitis E virus in experimentally infected pigs

The distribution of intravenously inoculated swine hepatitis E virus (HEV) was assessed by in situ hybridisation for a period of 50 days. Evidence of apparent clinical disease was found in only one pig in the HEV infected group. The only gross lesion observed was mildly enlarged mesenteric lymph nodes at 50 days post infection (dpi). Histopathologically, mild lymphoplasmacytic infiltration and focal hepatocellular necrotic lesions were found in HEV-infected pigs. Swine HEV nucleic acids were detected by RT-PCR in the faeces at 3 dpi in 100% of the 18 pigs infected with the virus. Thereafter, the number of positives declined.The most consistent and intense signal was found in the liver of infected animals using in situ hybridisation. The positive cells were hepatocytes, Kupffer cells, bile epithelial cells and interstitial lymphocytes. Swine HEV RNA was localised in the cytoplasm of the hepatocytes, with a slightly granular pattern of staining, but hybridisation signals were not observed in degenerative or vacuolated hepatocytes. HEV was much less frequently detected in extrahepatic tissues such as lymph nodes, tonsil, spleen and small and large intestine. It was concluded that swine HEV had replicated primarily in the hepatocytes and infection resulted in subclinical infection with minimal histopathological changes in the liver.     

猪血凝性脑脊髓炎病毒的血清学调查

应用血凝和血凝抑制试验检测了从辽宁省部分地区采集的猪血清中血凝性脑脊髓炎病毒（HEV）的抗体滴度。结果表明,368份血清样品中,有162份呈抗体阳性反应,阳性率高达44.0%。被采集血清的猪未表现出临床症状,说明该地区存在血凝性脑脊髓炎病毒的隐性感染。     

Pathomorphological and immunohistological findings in progeny of goats experimentally infected with pestiviruses.

A total of 25 pregnant goats without neutralizing antibodies against BVD virus were inoculated with two different pestivirus isolates at eight different stages of gestation. In both infection groups, various malformations were observed in fetuses and neonates. In three twins with neutralizing antibodies against BVD virus leukoencephalomalacia occurred, characterized by gelatinous transformation in the cerebral hemispheres. These lesions were comparable to alterations described in alternative pathology of Border disease in sheep. Although the immunohistological findings are characteristic for immunological tolerance and viral persistence, viable offspring persistently infected with pestivirus was not observed.     

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.