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Influence of isoprinosine on lymphocyte function in virus-infected feeder pigs
1989
Flaming, K.P. | Blecha, F. | Fedorka-Cray, P.J. | Anderson, G.A.
Pseudorabies is a porcine herpesvirus of major importance in the swine industry. Isoprinosine is an immunomodulating drug that has been shown to be beneficial in treating herpesvirus infections. Twenty-four 7-week-old pigs were allotted within litters to 1 of 4 groups: control, isoprinosine (ISO), pseudorabies virus (PRV), or isoprinosine and pseudorabies virus (ISO-PRV). Isoprinosine was administered daily for 16 days to the ISO and ISO-PRV groups (75 mg/kg of body weight/day, PO). Immunity in pigs in the PRV and ISO-PRV groups was challenged with pseudorabies virus (10(5) TCID50 units) on day 4. Rectal temperatures and viral excretion were monitored daily; total and differential leukocyte counts, lymphocyte response to mitogens, and interleukin-2 production were monitored every 4 days. Pigs challenge-inoculated with pseudorabies virus became ill, with the ISO-PRV group most severely affected. Rectal temperatures were high (P less than 0.05) in virally challenged pigs on days 5 to 12 and 14 to 16; isoprinosine did not alter this effect. Pseudorabies virus-infected pigs had leukocytosis (P less than 0.05) on days 12 and 16, primarily caused by neutrophilia. Concanavalin A-stimulated lymphocyte proliferation was decreased (P less than 0.06) in both PRV and ISO-PRV groups on day 12, compared with control pigs, but only in the PRV group on day 16. Pokeweed mitogen-stimulated lymphocyte proliferation was decreased (P less than 0.02) in ISO-PRV pigs on day 8 of the experiment. Interleukin-2 concentrations, pooled over all sampling days, were decreased (P less than 0.03) in pseudorabies virus-infected pigs. Viral excretion was not altered by isoprinosine treatment. These data suggest that pseudorabies virus infection decreased lymphocyte proliferative responses and interleukin-2 prodcution in pigs, and that isoprinosine did not mitigate these effects.
Show more [+] Less [-]Detection of turkey enteric coronavirus by enzyme-linked immunosorbent assay and differentiation from other coronaviruses
1989
Dea, S. | Tijssen, P.
A double-antibody ELISA for the detection of coronaviruses in intestinal contents from turkey poults with diarrhea was developed. Antibodies were raised in rabbits and guinea pigs against a Minnesota isolate of turkey enteric coronavirus (TCV) propagated in embryonating turkey eggs and were purified by density-gradient centrifugation. The specificity of antisera was confirmed by hemagglutination-inhibition and immunoelectron microscopy. Absorption of anti-TCV hyperimmune sera with egg extracts or egg ovalbumin and the use of different dilution and blocking buffers influenced the sensitivity and specificity of the ELISA. Reciprocal cross-reactivity was detected among turkey, chicken, bovine, and murine coronaviruses. Antisera to the transmissible gastroenteritis virus of swine, the rabbit enteric coronavirus, or the human coronavirus strain 299E failed to react with TCV. The TCV cross-related only moderately with the avian infectious bronchitis virus and the hemagglutinating encephalomyelitis virus of swine. Investigations with samples from 47 commercial turkey flocks in Quebec with episodes of transmissible enteritis revealed that the ELISA was more sensitive than was electron microscopy for dectection of TCV.
Show more [+] Less [-]Isolation and trypsin-enhanced propagation of turkey enteric (bluecomb) coronaviruses in a continuous human rectal adenocarcinoma cell line
1989
Dea, S. | Garzon, S. | Tijssen, P.
Turkey enteric coronavirus (TCV) from intestinal contents of diarrheal poults was isolated and serially propagated in HRT-18 cells, an established cell line derived from a human rectal adenocarcinoma. In these cells, TCV induced cytopathic changes, including polykaryocytosis, which depended on trypsin in the medium and incubation at 41 C. Viral antigens could be demonstrated in the cytoplasm by immunofluorescence, and extracellular virus was detected by an ELISA and negative electron microscopy. The cell-free virus had characteristics of TCV: shape, surface projections, buoyant density of 1.18 to 1.20 g/ml in sucrose, and hemagglutination of rat RBC. The one-step growth curve was complete by postinoculation hours 14 to 16, and maximal titers reached 9 to 9.5 log10 TCID50/ml during 5 passages, after which the titer remained stable. Electron microscopic examination of infected cell monolayers revealed budding of typical coronavirus particles through intracytoplasmic membranes and accumulation of complete virus in cytoplasmic vesicles. Late in the infection, aggregated progeny vial particles were detected near the outer surface of infected cells. One-day-old turkey poults inoculated orally with tissue culture-adapted TCV isolates developed mild to severe diarrhea.
Show more [+] Less [-]Effect of canine parvovirus on erythroid progenitors in phenylhydrazine-induced regenerative hemolytic anemia in dogs
1989
Brock, K.V. | Jones, J.B. | Shull, R.M. | Potgieter, L.N.D.
The effects of canine parvovirus (CPV) infection in dogs with hemolytic anemia was compared with the clinical effects of human parvovirus-induced aplastic anemia in human beings with chronic regenerative anemias. Phenylhydrazine was used to induce a transient, severe, hemolytic anemia in dogs to evaluate the effects of CPV infection on rapidly dividing bone marrow precursors. Erythrocyte colony-forming unit bone marrow cultures and cytologic examination of bone marrow were used to determine the effects of CPV infection on erythroid bone marrow precursors. The induced hemolytic anemia regenerated rapidly and although the bone marrow was infected, it was determined that CPV infection did not induce a detectable decrease in erythroid progenitors in dogs with severe hemolytic anemia.
Show more [+] Less [-]Hemagglutination and hemagglutination inhibition of turkey red blood cells with Mycoplasma hyopneumoniae
1989
Young, T.F. | Erickson, B.Z. | Ross, R.F. | Wannemuehler, Y.
The ability of Mycoplasma hyopneumoniae to agglutinate RBC was evaluated to develop an in vitro cytadsorption assay. Using swine RBC in a microtitration hemagglutination test, no agglutination or partial agglutination was detected. Comparison of RBC from various other species indicated that improved hemagglutination was obtained with RBC from turkeys. This hemagglutination was detected only when mycoplasma cells used in the assay had been frozen and thawed, heated at 50 C for 30 minutes, or treated with trypsin. Treatment of RBC with trypsin or neuraminidase enhanced hemagglutination. Possible surface lectin activity in M hyopneumoniae was evaluated by use of carbohydrates in a blocking assay; hemagglutination was not inhibited by any of 13 carbohydrates evaluated. Mycoplasma hyopneumoniae convalescent porcine serum and monoclonal antibodies against 2 M hyopneumoniae immunogens of molecular weights of 64,000 and 41,000 inhibited hemagglutination.
Show more [+] Less [-]Experimental rotavirus infection in three-week-old pigs
1989
Shaw, D.P. | Morehouse, L.G. | Solorzano, R.F.
Thirteen 3-week-old pigs that had been allowed to nurse for the first 16 to 18 hours after birth were orally inoculated with 1 X 10(6.5) TCID(50) of porcine rotavirus. All developed diarrhea, anorexia, and vomiting by postinoculation (PI) hour 30. These signs had abated by PI day 6. Villus blunting in the small intestine was most severe in the jejunum and ileum of pigs euthanatized between PI days 3 and 5. Villi had returned to nearly normal length by PI day 6, although fused villi were seen in a few locations in the distal portion of the jejunum and in the ileum. Virus was detected in the feces of inoculated pigs by isolation in cell cultures and by electron microscopy during the 7-day course of the experiment. There was 1 extraintestinal virus isolation from the lung of 1 pig at PI day 2. Infection and disease developed in the presence of serum-neutralizing antibody obtained by nursing seropositive sows. There was no significant change in neutralizing antibody titers in the 3-week-old pigs over the course of the experiment. In this experimental work, a model to study rotavirus infection in 3-week-old pigs has been developed.
Show more [+] Less [-]Rapid detection of Streptococcus suis serotype 2 in weaned pigs
1989
Moreau, A. | Higgins, R. | Bigras-Poulin, M. | Nadeau, M.
A survey to detect Streptococcus suis serotype 2 in 1,716 weaned pigs was done in Quebec. Forty-nine sow herds were included in this survey: in 26 herds, S suis serotype 2 had been isolated during the preceding 12 months and in 23 herds (control), the organism had not been detected during a previous study. Swab specimens of the nasal cavity and tonsils of pigs were obtained for bacteriological culture, and S suis serotype 2 was easily detected by the use of brain-heart infusion agar containing a Streptococcus-selective supplement and 5% goat antiserum raised against S suis serotype 2. After measurement of the diameter of the precipitation zone of 539 isolates, a slide agglutination test was performed to identify the S suis serot ype 2 isolates. The mean precipitation zone diameter obtained for group S suis serotype 2 was larger (P less than 0.001) than that for the group designated as "others." With slide agglutination test results as reference and on the basis of discriminant analysis to simulate detection of S suis serotype 2, 93.1% of all isolates were correctly classified, using the precipitation zone diameter as unique classification criterion. Relative specificity was 94.5% and relative sensitivity was 88.7%. Use of the precipitation zone diameter on a quantitative basis led to the proposal of a simple and reliable technique to screen swine herds for S suis serotype 2 in weaned pigs. Nasal and tonsillar swab specimens were obtained and analyzed concurrently for S suis serotype 2. The organism was found in both sites in only 20.4% of 103 carrier pigs. Nasal and tonsillar specimens yielded 55.3 and 65%, respectively, of all S suis serotype 2 isolates. Statistically significant difference was not observed between the numbers of S suis serotype 2 isolated from each site. Both sites permitted the recovery of S suis serotype 2 isolates; it was advantageous to use nasal and tonsillar swab specimens to determine the most reliable evaluation of S suis serotype-2 carrier status in weaned pigs.
Show more [+] Less [-]Sites of virus replication in the genital organs of boars inoculated in the cavum vaginale with pseudorabies virus
1989
Miry, C. | Pensaert, M.B.
To determine the sites of replication and the evolution of pseudorabies virus infection in boar genital organs, 5 Belgian Landrace boars were inoculated with pseudorabies virus unilaterally in the cavum vaginale of the testis. Virus replication took place only in cells of the tunica vaginalis of both cava vaginalia. Infection of the serosa led to exudative periorchitis and increased scrotal fluid, resulting in a severely swollen scrotal region. These experimental findings were similar to findings in boars with naturally aquired pseudorabies virus infection. Scrotal fluid contained large amounts of virus, making it a useful specimen for diagnosis of the disease in affected boars.
Show more [+] Less [-]Protection of mice against Brucella abortus infection by inoculation with monoclonal antibodies recognizing Brucella O-antigen
1989
Phillips, M. | Deyoe, B.L. | Canning, P.C.
Monoclonal antibodies recognizing the O-polysaccharide portion of Brucella abortus strain 2308 provided BALB/c mice with passive protection against challenge exposure with the homologous strain. Numbers of colony-forming organisms in the spleen were reduced by IgM and IgG monoclonal antibodies. Active immunization of mice, using B abortus 2308S lipopolysaccharide, resulted in production of IgM antibody at 14 days. Clearance of organisms in the actively immunized mice after challenge exposure at 14 days was nearly identical to that in passively immunized mice. Mice either passively or actively immunized were effectively protected from 0 to 28 days. Bacterial colonization of the spleen was observed to increase in both groups of mice at 56 days and indicated that humoral responses were effective in eliminating the organism in the early stages of infection, but other immune mechanisms were necessary for protection of mice in the later stage of infection with virulent strains of B abortus.
Show more [+] Less [-]Antigenic and restriction enzyme analysis of isolates of Campylobacter fetus subsp venerealis recovered from persistently infected cattle
1989
Wesley, I.V. | Bryner, J.H.
Thirty-two isolates of Campylobacter fetus subsp venerealis were obtained from 1 bull and 4 heifers with experimentally induced infection. When whole-cell antigens of isolates were cross titrated with antisera to the infecting strain, isolates from 3 heifers had limited antigenic variation, whereas whole-cell antigens of isolates from 2 cattle (the bull and a heifer) differed serologically from those of the infecting strain. Changes were detected specifically in 6 heat-labile antigens. Of the 6 heat-labile factors evaluated, all were initially present on the infecting parent strain, but not on early isolates obtained from 4 of the 5 cattle. Restriction enzyme analysis revealed minor variation in the DNA fingerprints of isolates obtained from individual cattle, thus implying stability of the Campylobacter genome once persistent infection is established. Isolates with identical restriction enzyme patterns expressed different heat-labile antigens. Correlation could not be found between the DNA electrophoretic pattern and the expression of heat-labile antigens.
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