Lactogenic antibody responses in cows vaccinated with recombinant bovine rotavirus-like particles (VLPs) of two serotypes or inactivated bovine rotavirus vaccines.

Triple-layered virus-like particles (VLPs) were produced in a baculovirus expression system from the two prevalent bovine rotavirus (BRV) serotypes, IND (P[5]G6) and 2292B (P[11]G10). Five groups of pregnant cows were inoculated intramuscularly and intramammarily with IND VLPs [BRV RF VP2, and IND VP4, 6, and 7, 250 microg per dose], 2292B VLPs [RF VP2, Cr VP4 (P[11]), and 2292B VP6 and 7, 250 microg per dose], combined IND/2292B VLPs (125 microg each VLP per dose), inactivated IND BRV (5x10(7)PFU per dose, pre-inactivation), or cell supernatant (mock-controls) in incomplete Freund's adjuvant. Serum, colostrum and milk were collected and tested for isotype-specific antibodies, and homologous and heterologous neutralizing antibodies (VN) to BRV by ELISA and VN tests, respectively. After vaccination, the IgG1 and homologous VN geometric mean antibody titers (GMTs) to BRV in serum of vaccinated groups were significantly (P<0.05) higher than in the mock-controls through postpartum day (PPD) 30. In colostrum, the IgG1 and IgA, and the homologous and heterologous VN GMTs of the IND VLP, 2292B VLP, combined IND/2292B VLP and the inactivated IND groups were significantly enhanced compared to the mock-controls, except for the heterologous VN GMTs in the inactivated IND group. However, the VLP vaccine groups had significantly higher homologous and heterologous VN GMTs than the inactivated IND group. The VN GMTs of the IND/2292B VLP group were statistically similar to the homologous VN GMTs of the IND or 2292B VLP groups, although the IgG1 GMT was lower. In milk, the IgG1 and homologous VN GMTs of the VLP groups were significantly higher than the inactivated IND or the mock-control groups through PPD30. However, the heterologous and homologous VN GMTs of inactivated IND group were statistically similar to the mock-control group at PPD0 and 30, respectively. These results demonstrate that the BRV antibody titers in serum, colostrum and milk are significantly enhanced by the use of triple-layered VLPs and inactivated IND vaccines, but significantly higher antibody responses were observed in the VLP vaccinated cows. The combined IND/2292B VLP vaccine induced comparable VN responses to BRV in serum, colostrum and milk compared to those induced by the individual IND or 2292B VLP vaccines, suggesting that at least two different serotypes can be mixed to confer maximum antibody responses to the incorporated serotypes.

First detection of bovine group B rotavirus in Japan and sequence of its VP7 gene.

An epizootic outbreak of diarrhea occurred in adult cows on a dairy farm in Hokkaido, Japan. One colostrum-fed calf inoculated with pooled feces of the 5 affected cows, developed mild diarrhea, and shed rotavirus-like particles which reacted with antiserum to group B rotavirus in immune electron microscopy. Cell culture immunofluorescence tests, RNA-polyacrylamide gel electrophoresis and RT-PCR confirmed that this virus was bovine group B rotavirus, which was designated the Nemuro strain. Additional 2 colostrum-deprived calves inoculated with feces of the first calf also developed diarrhea and shed virus, suggesting that this group B rotavirus might be the etiological agent of the outbreak of adult cow diarrhea. The identities of the nucleotide (nt) and deduced amino acid (aa) sequences of the Nemuro VP7 gene were high (93-95% in nt and 96-97% in aa) and low (61-63% in nt and 49-61% in aa) compared to those of the published corresponding genes from 3 bovine and 2 other mammalian (human and rat) strains of group B rotaviruses, respectively. To our knowledge, this is the first report on the presence of bovine group B rotavirus in Japan.

Passive immunity to bovine rotavirus in newborn calves fed colostrum supplements from cows immunized with recombinant SA11 rotavirus core-like particle (CLP) or virus-like particle (VLP) vaccines.

Heterotypic passive immunity to IND (P/5/G6) bovine rotavirus (BRV) was evaluated. Three groups of calves (n = 5 per group) were fed 1% pooled colostrum supplements (birth to 7 days of age) from BRV seropositive cows vaccinated with recombinant SA11(P/2/G3) rotavirus-like particles (VLPs), recombinant SA11 rotavirus core-like particles (CLPs), or inactivated SA11 rotavirus (SA11). Control calves (n = 5 per group) received either pooled colostrum from unvaccinated (BRV field exposure seropositive) control cows, or no colostrum. IgG1 antibody titers to IND BRV for the pooled colostrum were: 1,048,576 (VLP); 1,048,576 (CLP); 262,144 (SA11); and 16,384 (control colostrum). Elevated titers of BRV neutralizing (VN) antibodies were present in VLP colostrum (98,000), and SA11 colostrum (25,000), but not in CLP colostrum (1400), compared to colostrum from nonvaccinates (2081). Calves were orally inoculated with virulent IND BRV at 2 days of age and challenged at post-inoculation day (PID) 21. Calves were monitored daily for diarrhea and faecal BRV shedding through PID 10 and post-challenge day (PCD) 10. After colostrum feeding, the IgG1 antibody titers were highest in serum and faeces of calves fed VLP and CLP colostrum, but VN and IgA antibodies were highest in calves fed VLP colostrum. After BRV inoculation, calves fed colostrum from vaccinated cows had significantly fewer days of BRV-associated diarrhea and BRV shedding than control calves. All calves fed VLP colostrum were protected from diarrhea after BRV inoculation; two calves shed BRV. In the CLP colostrum group, one calf developed BRV-associated diarrhea and all calves shed virus. In the SA11 colostrum group, three calves developed BRV-associated diarrhea and four calves shed virus. BRV-associated diarrhea and shedding occurred in 9 of 10 control calves. Active IgM antibody responses occurred in faeces and/or serum of most calves after BRV inoculation. However, the highest active antibody responses (IgM and IgG1 in serum, and IgM, IgG1 or IgA in faeces) after BRV inoculation were in calves fed control or no colostrum, in association with clinical diarrhea in most of these calves. After challenge at PID 21, BRV-associated diarrhea and shedding were of short duration or absent, in all groups. These results demonstrate the efficacy of colostrum from VLP vaccinated cows to provide heterologous, passive protection against BRV diarrhea and shedding in calves. In comparison, calves fed CLP or SA11 colostrum were only partially protected against BRV diarrhea or shedding.

Active and passive immune responses to transmissible gastroenteritis virus (TGEV) in swine inoculated with recombinant baculovirus-expressed TGEV spike glycoprotein vaccines.

OBJECTIVE: Baculovirus-expressed transmissible gastroenteritis virus (TGEV) spike (S) glycoprotein vaccines were inoculated parenterally in swine to determine whether such vaccines could induce serum and whey virus-neutralizing (VN) antibodies and protective lactogenic immunity for TGEV-challenge-exposed pigs. ANIMALS AND PROCEDURES: 3 recombinant baculoviruses that expressed full or partial length TGEV Miller strain S glycoproteins were inoculated SC in 17 conventionally raised 11-day-old TGEV-seronegative pigs to determine whether the recombinant S glycoproteins would elicit serum VN antibodies. Eleven TGEV-seronegative pregnant sows were inoculated SC or intramammarily with subunit vaccines (R2-2 or R3-5) or control proteins. Pigs born to 9 of the 11 sows were challenge exposed at 4 to 5 days of age with the virulent Miller strain, and passive immunity was assessed. Serum and whey antibody responses to TGEV were analyzed by VN and ELISA testing. RESULTS: Recombinant S glycoproteins (R2-2 or R3-5) containing the 4 major antigenic sites induced similar VN antibody titers to TGEV in serum and colostrum, but low (some sows) or no VN antibody titer was detected in milk. Subcutaneous inoculation of sows with R2-2 or R3-5 elicited IgG, but not IgA antibodies to TGEV in colostrum. Morbidity was 100%, and mortality ranged from 20 to 80% in TGEV challenge-exposed pigs nursing sows inoculated SC or intramammarily with TGEV S glycoprotein vaccines. CONCLUSIONS AND CLINICAL RELEVANCE: Parenterally administered TGEV S glycoprotein vaccines elicit VN antibodies to TGEV in serum and colostrum that do not fully provide active or passive immunity in swine.

Isotype-specific antibody responses to rotavirus and virus proteins in cows inoculated with subunit vaccines composed of recombinant SA11 rotavirus core-like particles (CLP) or virus-like particles (VLP).

The isotype antibody responses to bovine IND P5, G6 and simian SA11 P2, G3 rotavirus and SA11 rotavirus proteins (VP4, VP6 and VP7) in serum, colostrum and milk were analysed by ELISA in three groups of vaccinated cows and nonvaccinated controls. Pregnant cows were vaccinated intramuscularly and intramammarily with recombinant baculovirus-expressed SA11 rotavirus VLP (triple-layered virus-like particles containing rotavirus VP2, VP4, VP6 and VP7); CLP (double-layered core-like particles containing rotavirus VP2 and VP6); or inactivated SA11 rotavirus, respectively. Rotavirus antigen titers were highest (30-200-fold) in ELISA in the VLP vaccine compared to the inactivated SA11 vaccine. The IgG1, IgG2 and IgM geometric mean antibody titers (GMT) to rotavirus (titers to bovine rotavirus vs SA11 rotavirus did not differ significantly for any isotype or group) and the IgG2 GMT to VP6 in serum at calving in the vaccinated groups were significantly (P < 0.05) higher than in the control group. In colostrum, IgG1 and IgA rotavirus antibody titers were significantly elevated for VLP (IgG1 GMT 832225; IgA GMT 16384), CLP (IgG1 GMT 660561; IgA GMT 10321) and SA11 (IgG1 GMT 131072; IgA GMT 1448) vaccinated cows compared to control cows (IgG1 GMT 11585; IgA GMT 45). The IgG1 and IgA GMT to rotavirus were significantly elevated (6-100-fold) in milk of VLP and CLP vaccinated cows compared to SA11 vaccinated or control cows. The isotype antibody responses to VP6 in serum, colostrum and milk paralleled the responses to rotavirus, but titers were approximately 2-10-fold lower. Only cows vaccinated with VLP had significantly enhanced serum, colostral and milk antibody titers to rotavirus VP4 and VP7. These results demonstrate that rotavirus antibody titers in serum, colostrum and milk are significantly enhanced by use of non-infectious VLP, CLP and inactivated SA11 rotavirus vaccines, but the VLP or CLP vaccines induced the highest antibody responses, corresponding to their higher rotavirus antigen titers measured by ELISA.

Contribution of passive immunity to porcine respiratory coronavirus to protection against transmissible gastroenteritis virus challenge exposure in suckling pigs.

OBJECTIVE: To determine the ability of porcine respiratory coronavirus (PRCV) infections to induce passive immunity in suckling pigs to transmissible gastroenteritis virus (TGEV) challenge exposure. DESIGN AND ANIMALS: 4 TGEV seronegative sows and their litters (group A) served as controls, whereas 2 other groups (B and C) of sows (also TGEV seronegative) were oronasally inoculated with live PRCV during 1 or 2 subsequent pregnancies, respectively. PROCEDURE: Effectiveness of passive immunity provided to pigs via colostrum and milk was assessed after TGEV challenge exposure, and TGEV antibody responses in colostrum and milk were analyzed. RESULTS: Mortality in the 3 groups of young pigs correlated with severity of clinical signs of TGEV infection and was highest in control litters (86% in group-A pigs) and lowest in litters of sows inoculated with PRCV in 2 subsequent pregnancies (14% in group-C pigs). Virus-neutralization and IgA and IgG TGEV antibody titers of milk collected from sows at challenge exposure had significant positive correlation with litter survival. Significantly higher numbers of TGEV-specific IgA and IgG antibody-secreting cells were found in group-A pigs than in group-C pigs, suggesting that high titer of maternal antibodies (induced in group-C sows multiply exposed to PRCV) may interfere with active antibody responses. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that, in PRCV-infected pig herds, multiple exposures of pregnant sows are associated with higher IgA and IgG antibody titers to TGEV in milk, and these titers contribute to protection against TGEV infection.

Infection and cross-protection studies of winter dysentery and calf diarrhea bovine coronavirus strains in colostrum-deprived and gnotobiotic calves.

OBJECTIVE: To investigate in vitro antigenic relations, in vivo cross-protection, and isotype antibody responses to a winter dysentery (WD) and calf diarrhea strain of bovine coronavirus (BCV). DESIGN AND ANIMALS: Gnotobiotic and colostrum-deprived calves were inoculated oronasally with a WD (DBA) or a calf diarrhea (DB2) BCV, and were challenge exposed with the heterologous BCV. PROCEDURE: Nasal swab and feces specimens and blood samples were collected. Fecal and nasal specimens were assayed for BCV shedding by antigen-capture ELISA or immune electron microscopy. Bovine coronavirus antigens were detected in nasal epithelial cells by immunofluorescence. Antibody titers to BCV in serum were assayed by virus neutralization (VN), and BCV antibody isotype titers in feces and sera were quantitated by ELISA. RESULTS: All calves developed diarrhea and shed BCV nasally and in feces, then recovered and were protected from BCV-associated diarrhea after challenge exposure with the heterologous BCV. After challenge exposure with either strain, fecal shedding of DBA was detected in 1 of 4 calves and nasal shedding of DB2 was detected in 2 of 4 calves. Immunoglobulin M was the principal coproantibody to BCV early, followed predominantly by IgA. Immunoglobulin G1 coproantibody titers to BCV were low, but increased after challenge exposure. Immunoglobulin G1 antibodies were predominant in serum. After challenge exposure, all serum antibody isotype titers increased except IgG2. The VN antibody responses paralleled serum IgG1 antibody responses. CONCLUSIONS AND CLINICAL RELEVANCE: Immunoglobulin A coproantibodies at challenge exposure were associated with protection against diarrhea. Nasal shedding of BCV after challenge exposure confirmed field data documenting reinfection of the respiratory tract of cattle, suggesting that, in closed herds, respiratory tract infections constitute a source of BCV transmission to cows (WD) or young calves.

Prevention of human rotavirus-induced diarrhea in gnotobiotic piglets using bovine antibody.

The efficacy of passively administered bovine antibody for preventing human rotavirus (HRV)-induced diarrhea was investigated using a gnotobiotic pig model. Cows were immunized with inactivated HRV serotypes 1 (Wa) and 2 (S2) and simian rotavirus serotype 3 (SA11), and immune colostrum and milk were collected. Antibody concentrates derived from these materials were fed to germ-free piglets that were subsequently inoculated with HRV Wa. Both viral shedding and diarrhea were effectively reduced or eliminated in a dose-dependent manner as a result of HRV immune antibody feeding. A quantitative virus-neutralizing (VN) antibody method permitted assessment of the functional antibody dose required to achieve a 50% reduction of disease (PD50). PD50 dose levels of 15.8 and 19.5 x 10(6) VN antibody units were determined for inhibition of diarrhea and viral shedding, respectively. Studies reported here provide new information on the quantitative relationship between protective antibody dose and diarrheal disease response.

Epidemiologic factors and isotype-specific antibody responses in serum and mucosal secretions of dairy calves with bovine coronavirus respiratory tract and enteric tract infections.

Blood, feces, and nasal swabs specimens were collected 12 to 24 hours after birth and then 3 times/week (blood only once per week) from one group of 10 calves until they were 10 weeks old and from a second group of 10 calves until they were 10 to 20 weeks old. Colostrum was collected from all calves' dams and tears from 5 randomly selected calves in the first group. All fecal and nasal specimens were assayed for bovine coronavirus (BCV) antigens by ELISA. Nasal epithelial cells were examined for BCV antigens by direct immunofluorescence. Isotype antibody titers to BCV in all samples from 5 calves in group 1 were evaluated by ELISA. Zinc sulfate turbidity (ZST) values were determined on the first serum samples taken from all calves in group 1. To determine whether any correlation existed between ZST values, isotype antibody titers to BCV (12 to 24 hours after birth), number of respiratory sick days, number of enteric sick days, or days to first shedding of virus, a Spearman rank order correlation coefficient was done. Bovine coronavirus respiratory tract and enteric tract infections were common on this farm. Most initial infections developed when calves were 1 to 3 weeks old; however, there were also multiple incidences of shedding of viral antigens or seroconversions at later times during the study. Persistence of infection or reinfection of the upper respiratory tract with BCV was common. Colostral antibody titers to BCV (IgG1) were in all cows at moderate amounts; however, calf serum antibody titers and ZST values (12 to 24 hours after birth) were highly variable.(ABSTRACT TRUNCATED AT 250 WORDS)

Mucosal and systemic antibody responses to bovine coronavirus structural proteins in experimentally challenge-exposed calves fed low or high amounts of colostral antibodies.

Ten colostrum-deprived calves were assigned to 1 of 2 treatment groups (5 calves/group), and fed colostrum that had either low (naturally infected cows) or high (immunized cows) antibody titers to bovine coronavirus (BCV). All calves were inoculated orally and intranasally with virulent BCV when they were 24 to 48 hours old and challenge exposed 21 days later. Blood, feces, nasal secretions, tears, saliva, and bronchoalveolar lavage (BAL) fluids were collected weekly from each calf for 5 weeks after inoculation. The titers to whole BCV or the relative amounts of isotype-specific antibodies to BCV structural proteins were evaluated in these samples by ELISA or immunoblotting, respectively. Both pools of colostrum contained primarily IgG1, IgG2, and IgA antibodies to the E2 and E3 BCV proteins. Calves fed the high-titer colostrum had correspondingly higher amounts of passive IgG1 and IgA antibodies to whole BCV and to the E2 and E3 BCV proteins in serum, feces, and BAL fluid at postinoculation week 1 than those calves fed low-titer colostrum. Active IgG1, IgA, and IgM antibody responses in serum and active IgA and IgM antibody responses in most mucosal secretions to whole BCV and to the E2 and E3 proteins were lower or delayed in calves fed high-titer colostrum, compared with responses in calves fed low-titer colostrum. In contrast, increased responses to the BCV N protein were observed in all samples (except in serum and BAL fluid) in the calves fed high-titer colostrum, compared with calves fed low-titer colostrum. Upon challenge exposure, responses to E2 and E3 BCV proteins in serum and BAL fluid were lower in the group fed high-titer colostrum, compared with those in the group fed low-titer colostrum. Our findings indicate that the level of passive immunity in calves at the time of BCV inoculation can influence the development of active antibody responses in serum, feces, and mucosal secretions to whole BCV and to some BCV proteins individually.

Experimentally induced coronavirus infections in calves: viral replication in the respiratory and intestinal tracts.

Eleven 3- to 50-day-old colostrum-deprived gnotobiotic calves and seven 25- to 63-day-old colostrum-deprived conventional calves were allotted into 3 groups. Each group was inoculated with a fecal isolate of bovine coronavirus via different routes: orally/intranasally OR/IN, No. 1 through 8, group 1 calves; OR, No. 9 through 13, group 2 calves; IN, No. 14 through 18, group 3 calves. Nasal swab specimens and fecal specimens were collected daily and were examined for coronavirus antigen by use of direct immunofluorescent staining (nasal epithelial cells) or by use of immune electron microscopy (fecal specimens). All but 4 calves (No. 11, 13, 17, and 18) were euthanatized on postinoculation days (PID) 3 to 7. Calves 11 and 17 became severely dehydrated and died at PID 5. Calves 13 and 18 were evaluated for nasal and fecal shedding of coronavirus through PID 14. Distribution of coronavirus antigen in the respiratory and intestinal tracts of the 14 euthanatized calves was evaluated by use of direct immunofluorescent staining. All calves developed profuse diarrhea by PID 2 to 4; however, calves did not develop clinical signs of respiratory tract disease before euthanasia or death. Inoculated calves shed coronavirus in their feces as detected by use of immune electron microscopy. Infected nasal epithelial cells were detected in all but 2 orally inoculated calves (No. 9 and 10). Route of inoculation influenced the sequence of initial detection of coronavirus antigen from fecal specimens or nasal swab specimens.(ABSTRACT TRUNCATED AT 250 WORDS)

Enteric viral infections of calves and passive immunity.

At least eight viruses have been identified, four within the last 5 yr, that produce diarrhea and pathological intestinal lesions in experimentally inoculated calves. Coronavirus and rotavirus frequently are associated with the neonatal calf diarrhea syndrome, but the etiologic role of the newly identified viruses is undefined. All diarrheal viruses replicate within small intestinal epithelial cells, resulting in variable degrees of villous atrophy. Immunity against these viral infections, therefore, must be directed toward protection of the susceptible intestinal epithelial cells. Because most of these viral infections occur in calves less than 3 wk of age, passive lactogenic immunity within the gut lumen plays an important role in protection. This report reviews methods of boosting rotavirus antibody responses in bovine mammary secretions and analyses of passive and active immunity in calves supplemented with colostrum and challenged by rotavirus. Results indicate rotavirus immunoglobulin G1 antibodies in colostrum and milk were elevated after intramuscular and intramammary vaccination of pregnant cows with an Ohio Agricultural Research and Development Center rotavirus vaccine but not after intramuscular immunization with a commercial rota-coronavirus vaccine. Feeding colostrum from intramuscular plus intramammary immunized cows to newborn calves challenged by rotavirus prevented diarrhea and shedding of rotavirus.

Passive immunity to bovine rotavirus in newborn calves fed colostrum supplements from immunized or nonimmunized cows.

Colostrum was collected and pooled from each of five cows in three experimental groups: group I cows received intramuscular and intramammary inoculations of adjuvanted modified live Ohio Agricultural Research and Development Center rotavirus vaccine; group II cows were injected intramuscularly with a commercial modified-live rota-coronavirus vaccine; and group III cows were uninoculated controls. Pooled colostrum from group I cows had higher (P less than 0.05) enzyme-linked immunosorbent assay (ELISA) immunoglobulin G (IgG1) and virus neutralization (VN) rotavirus antibody titers (ELISA IgG1 = 2,413,682; VN = 360,205) than did colostrum from group II (ELISA IgG1 = 8,192; VN = 4,395) or group III cows (ELISA IgG1 = 5,916; VN = 2,865). The antibody titers of these last two colostrum pools did not differ (P greater than 0.05). Samples of these colostrum pools were fed as daily supplements (percent [vol/vol] in cow's milk infant formula) to 28 newborn, unsuckled, antibody-seronegative, male Holstein calves. Eight calves received no supplemental colostrum. The calves were orally challenged with virulent bovine rotavirus and monitored daily for diarrhea and fecal rotavirus shedding. Diarrhea and rotavirus shedding occurred in the eight calves fed no supplemental colostrum and persisted longest in this group. The pooled colostrum from group I cows protected eight of eight calves from both rotavirus diarrhea and shedding when fed as a 1% supplement. The pooled colostrum from neither group II nor group III cows protected 12 other calves against rotavirus diarrhea or shedding when fed at the same concentration (1%). Six rotavirus-challenged calves fed 0.1% supplemental colostrum from group I cows and two calves fed 10 and 50% supplemental colostrum from control cows displayed partial passive immunity, exemplified by delayed onset and shortened duration of rotavirus-associated diarrhea and virus shedding.

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.

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.

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.