PubMed Health⌕ Search

Biomedical subjects

E H Bohl

Publications and source records attributed to E H Bohl.

At least 37 records · Page 2Linked to original sources

Immune electron microscopy of transmissible gastroenteritis virus and rotavirus (reovirus-like agent) of swine.

Immune electron microscopy (IEM) was developed as a diagnostic aid for detecting and identifying transmissible gastroenteritis virus and rotavirus (reovirus-like agent) in fecal and intestinal contents from cases of gastroenteritis in young pigs. Variables involved in use of direct IEM and its sensitivity were determined. Aggregates of virus coated with specific antibody were seen in virus samples mixed with homologous convalescent antiserum, but not in control samples containing preexposure serum or antibody directed against a heterologous virus. At least a ten fold enhancement of the sensitivity of direct IEM for virus detection was accomplished using indirect IEM employing rabbit anti-porcine IgG to further aggregate virus-antibody complexes. The technique was used to investigate the size and morphology of the porcine rotavirus. Particles ranged from 55 to 70 nm in diameter and had capsomere structures. Morphologically, the porcine rotavirus resembled the calf and human rotaviruses. By IEM, employing specific antiserums for each virus, porcine rotavirus was found to be antigenically related to these 2 viruses, but not to the reovirus type 3.

Animals↗

Cell culture propagation of porcine rotavirus (reovirus-like agent).

Two isolates of porcine rotavirus (reovirus-like agent) were isolated and passaged in primary procine kidney cell cultures. Viral infectivity for cells was monitored by immunofluorescence because viral cytopathic effect was moderate. Successful passage of virus in cell culture required that viral suspensions obtained from infected cell cultures be treated with pancreatin prior to inoculation onto cell monolayers. Porcine rotavirus passage in cell culture also was accomplished, using trypsin treatments in lieu of pancreatin treatments. Porcine rotavirus passaged 10 times in cell culture infected gnotobiotic pigs and caused diarrhea. Gnotobiotic pigs that recovered from this infection were resistant to challenge exposure with porcine rotavirus but were susceptible to challenge exposure with transmissible gastroenteritis virus. As determined by immunofluorescent cross reactions, porcine rotavirus was found to be antigenically related to the human and bovine rotaviruses but not to reovirus type 3 or to transmissible gastroenteritis virus.

Animals↗

Pathogenicity of an attenuated strain of transmissible gastroenteritis virus for newborn pigs.

The pathogenicity of a cell culture-attenuated strain of transmissible gastroenteritis virus for newborn pigs was investigated. Newborn (1- to 2-day-old) pigs were orally given 2 x 10(6) plaque-forming units of attenuated virus. All pigs developed mild diarrhea, but deaths did not occur. As determined by immunofluorescence and villous atropy, infection of the small intestine was limited to the caudal 50 to 66%. Fluorescing cells and atrophic villi were seen from 2 to 3 days until 6 to 7 days after exposure. Attenuated virus-exposed pigs produced circulating virus-neutralizing antibodies detectable as early as 5 days after exposure. By contrast, all pigs orally given 1 x 10(2) pig infective doses of virulent transmissible gastroenteritis virus developed severe diarrhea, and almost all of those not killed died within 2 to 5 days after exposure. In the latter pigs, the entire length of the small intestine, except for the first 4 to 5 cm, was infected with virus by 24 to 36 hours after exposure.

Alkaline Phosphatase↗

Local and systemic cell-mediated immunity against transmissible gastroenteritis, an intestinal viral infection of swine.

The occurrence of local and systemic cell-mediated immunity (CMI) in swine against transmissible gastroenteritis (TGE) virus, a coronavirus that infects the epithelial cells of the small intestine of swine, has been investigated. Leukocyte suspensions prepared from intestinal lamina propria (LP) and spleen were each incubated with and without inactivated TGE virus. The supernatants from these cultures were tested for the presence of macrophage migration inhibition factor (MIF) by the indirect MIF test using guinea pig peritoneal exudate cells as the source of macrophages. All swine serologically negative for TGE antibodies (i.e., unexposed to TGE virus) were also negative for MIF production by lymphocytes from both sources. In 10 of 10 animals given TGE virus subcutaneously, MIF production by splenic lymphocytes was significantly greater (p less than or equal to 0.01) than by LP lymphocytes, and in 9 of 10 of these animals MIF production by LP lymphocytes was negligible (percentage of inhibition of migration less than 3%).

Animals↗

Passive immunity in transmissible gastroenteritis of swine: immunoglobulin characteristics of antibodies in milk after inoculating virus by different routes.

Pregnant swine were exposed to transmissible gastroenteritis (TGE) virus by different routes, and their serum, colostrum, and mild were examined for titer and immunoglobulin (Ig) class of antibodies. When 2 to 4 days old, the litters of most of these animals were challenged with virulent TGE virus to determine the effectiveness of passive immunity. After two oral/intranasal exposures to attenuated virus, none of the six pregnant animals became sick. TGE antibodies in milk were primarily or solely of the IgG class, although low levels of IgA antibodies were detected in three animals. Pigs in the five challenged litters received some passive immunity, the mortality being 25%. After intramuscular injection of six pregnant swine with virulent virus, two types of clinical and immunological responses were observed, presumably dependent on whether the gut was infected by an hematogenous spread of the virus. Three became sick, showing typical clinical signs of TGE, and their immunological response was characterized by the occurrence in milk of antibodies of the IgA class. A good degree (0% mortality) of passive immunity occurred upon challenge of the suckling pigs. In contrast, in three pregnant animals that did not sicken, antibody in milk was primarily of the IgG calss, and poor (69% mortality) passive immunity occurred. After intramammary injections of three pregnant swine with virulent virus, no sickness was observed and the immunological response was characterized by the occurrence in colostrum of high titers of TGE antibodies that were primarily or solely of the IgG class; good (0% mortality) passive immunity occured. The occurrence in milk of TGE antibodies of the IgA class was associated with an intestinal infection, whereas antibodies of the IgG class resulted from a parenteral antigenic stimulation. The role of antigenic stimulation of the intestinal tract for providing antibodies in milk of the IgA class is discussed. Passive immunity against intestinal infection with TGE virus was generally more complete in pigs ingesting antibodies of the IgA than of the IgG class.

Absorption↗

Protection against neonatal enteric colibacillosis in pigs suckling orally vaccinated sows.

Pregnant swine were vaccinated with 1 of 2 enteropathogenic strains of Escherichia coli, and their pigs were challenge inoculated with the homologous strain 1.5 hours after the entire litter was born. Seventeen sows were vaccinated orally by feeding viable cultures on 3 consecutive days, 6 sows were given 2 intramuscular (IM) injections of viable cultures at 10- to 14-day intervals, 6 sows were given 2 IM injections fo formolized cultures at 10- to 14-day intervals, and 12 sows were not vaccinated. The pigs from sows which had been orally vaccinated with viable cultures were protected during the 10-day observation period against diarrhea, as well as against death, when the newborn pigs were challenge inoculated with the homologous strain. Most of the pigs from sows which had been vaccinated IM were protected against death, but few neonates were protected against the diarrheal effects of challenge exposure with the homologous strain. Challenge-inoculated pigs suckling nonvaccinated sows had diarrhea and became dehydrated, and many died. Fewer viable E coli were usually recovered from the homogenized intestinal contents or intestinal segments of newborn pigs which did not have diarrhea than from similar specimens of diarrheal pigs. Microscopic examination of segments of the small intestine revealed that large numbers of E coli were closely associated with the ileal mucosa of newborn pigs killed in the acute phase of neonatal enteric colibacillosis caused by either strain, but organisms were not detected in this location in the pigs which remained normal after challenge exposure. It is concluded that swine which have been vaccinated by feeding large numbers of viable E coli late in gestation can provide good protection to their suckling pigs against the effects of challenge inoculation with the homologous enteropathogenic strain.

Administration, Oral↗

Passive immunity in transmissible gastroenteritis of swine: intramuscular injection of pregnant swine with a modified live-virus vaccine.

Nine pregnant swine were intramuscularly injected with a modified live-virus transmissible gastroenteritis (TGE) vaccine at approximately 6 and 2 weeks before parturition. The TGE antibody levels in milk whey rapidly decreased within a few days after parturition, and the antibodies were mainly, if not solely, of the immunoglobulin G (IgG) class. Nursing newborn pigs were challenge exposed with virulent TGE virus when 2 to 4 days old. All pigs in 8 of the 9 litters became affected with typical signs of TGE. Of the pigs nursing vaccinated sows, 55% died, and of the 9 vaccinated sows, 8 became affected after challenge exposure of their litters. Subsequently, the antibody titers in serum and milk markedly increased in all 9 sows, with the antibody in milk being predominantly of the immunoglobulin A (IgA) class. Serum samples of prenursing newborn pigs from vaccinated sows did not contain detectable TGE antibodies. The capability of the vaccinated sows to provide only limited passive immunity to their suckling pigs was considered to be due mainly to the low titer and class (IgG) of TGE antibody existing in the milk of the sows at the time the newborn pigs were challenge exposed.

Animals↗

Porcine parvovirus: natural and experimental infections of the porcine fetus and prevalence in mature swine.

Antibodies against porcine parvovirus were detected in 17 of 116 prenursing pig sera. Antibodies against transmissible gastroenteritis or ECPO-6 (an enterovirus) were not detected in prenursing sera of the pigs tested. Seventy-seven percent of 129 serum samples from 23 Ohio farms and 82% of 96 samples from slaughter plants in Ohio were serologically positive for porcine parvovirus. Mummies or other abnormalities were not observed in newly born pigs exposed to porcine parvovirus by the transuterine route 101 days after gestation. Indirect evidence suggested that the virus had not spread to other fet uses following exposure after 101 days at least not in a sufficient amount of time to stimulate detectable antibody. Direct intrafetal exposure to porcine parvovirus (i.m. injection, transutero) after 62 days of gestation resulted in dealth and mummification of the two fetuses, and apparently in the subsequent spread of the virus, as five of nine live pigs born were serologically positive for porcine parvovirus and these five pigs had not been injected with the virus. Immunoglobulin G was detected in all newborn pigs irregardless of known antigenic stimulation or the presence of specific antibody. In general, the presence of immunoglobulin M or immunoglobulin A in fetal serum was correlated with a history of antigenic stimulation or the presence of detectable antibody.

Animals↗

Antibody responses in serum, colostrum, and milk of swine after infection or vaccination with transmissible gastroenteritis virus.

The antibody response of pregnant swine to transmissible gastroenteritis (TGE) virus was studied, with special reference to the titers and the immunoglobulin (Ig) class of TGE neutralizing antibodies in colostrum and milk. Animals vaccinated twice intramuscularly or intramammarily with live attenuated TGE virus developed high levels of antibodies in serum and colostrum, but the levels in milk declined markedly within a few days post-farrowing. In contrast, animals naturally or experimentally infected with virulent virus generally developed lower levels of antibodies in serum and colostrum but maintained higher levels in milk, as compared to the vaccinated animals. Gel filtration studies indicated that antibodies in milk from vaccinated animals were primarily of the IgG class, whereas those from the naturally or experimentally infected animals were primarily of the IgA class. The ability of sows to transmit a high degree of passive immunity to their suckling progeny was more closely associated with TGE antibodies of the IgA than the IgG class. Present evidence suggests that high levels of TGE antibodies of the IgA class occur in milk as a result of an infection of the intestinal tract. Probable reasons for this are discussed.

Animals↗

Isolation of porcine immunoglobulins and determination of the immunoglobulin classes of transmissible gastroenteritis viral antibodies.

The porcine immunoglobulins M (IgM), A (IgA), and G (IgG) were isolated and purified and some of the properties of the porcine milk IgA were examined. Monospecific antisera which were prepared against these immunoglobulins in rabbits were then used to absorb a particular class of immunoglobulin from sow serum, colostrum, and milk in an attempt to identify the immunoglobulin classes of neutralizing antibodies to the porcine enteric virus, transmissible gastroenteritis (TGE). The results of these absorption studies suggest that in colostrum and milk from sows experimentally (orally) or naturally infected with live virulent TGE virus, IgA is the predominant immunoglobulin class of TGE antibodies. Both IgA and IgG TGE antibodies appeared to be present in the serum from these sows, but with IgG TGE antibodies predominating. In contrast, in the serum, colostrum and milk from sows vaccinated intramuscularly or intramammarily with live attenuated TGE virus, the TGE antibody activity was associated mainly with the IgG class of immunoglobulins. These results provide additional data indicating that the route of infection or vaccination markedly influences the immunoglobulin class of antibodies in colostrum and milk. Secondly, IgA antibodies in mammary secretions are probably essential for providing optimal passive immunity of nursing pigs against infection with TGE virus.

Adsorption↗

The pattern of enteroviral infections in a herd of swine.

Twenty-six pigs from four litters in a healthy herd of swine were examined periodically for the fecal excretion of viruses by the use of porcine kidney cell cultures. Viruses were initially isolated from all pigs between 34 to 64 days of age. The pigs within each litter began shedding virus in their feces approximately at the same time, usually within one week, and the type of virus initially recovered was usually the same. Subsequently, waves of infection with different enteroviruses appeared to occur during the observation period of six months. At least six antigenically different viruses were isolated from this herd over a 26-month period. Most, if not all, of these viruses were considered to belong to the enterovirus group. No disease was associated with these enteroviral infections. The colostrum and milk of sows contained significant amounts of enteroviral antibodies. Prior to nursing, the serum of new-born pig contained no enteroviral antibodies but, shortly after nursing, high titers of such antibodies were present in the serum. Antibodies were detected in the feces of suckling pigs.

Animals↗