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Biomedical subjects

H S Joo

Publications and source records attributed to H S Joo.

At least 37 records · Page 2Linked to original sources

Improved performance of a large pig complex after sequential nursery depopulation.

An attempt was made to compare the productivity and financial benefits of nursery depopulation on five large pig farms which were all part of one complex. Each farm had been experiencing poor post weaning performance for 12 months, and had previously been infected with porcine reproductive and respiratory syndrome virus (PRRSV). A plan to depopulate each nursery sequentially was established, and the pigs were moved to fattening facilities on one of the farms (farm 3) where space was available. Over a four week period, the nurseries of farms 1, 2, 4 and 5 were emptied, cleaned and disinfected, and any changes in nursery performance, mortality and the seroprevalence of antibodies to PRRSV were then assessed for one year. The financial benefit to the entire farm complex was analysed by using partial budget methods. During the year a net benefit of $1,708,431 was assessed to the farm complex owing to the increased numbers of marketable pigs and the reduced antibiotic costs. There were highly significant improvements in nursery growth rate and decreases in mortality on farms 1, 2, 4 and 5, and antibodies to PRRSV were detected on farms 3 and 4 but not on farms 1, 2 and 5. The inability to empty the farm 3 fattening facility, which housed the pigs from the other sites, may have led to the maintenance of its PRRSV positive status and could have served as the source of virus for farm 4.

Animal Husbandry↗

Pathogenesis of plaque variants of porcine reproductive and respiratory syndrome virus in pregnant sows.

OBJECTIVE: To determine whether porcine reproductive and respiratory syndrome virus plaque variants vary in their pathogenicity in causing late-term reproductive failure. DESIGN: Four groups of 2 sows each at 86 days of gestation were inoculated intranasally with PRRSV small (MN-Hs) and large (MN-HI) plaque variants, field isolate, and cell culture medium, respectively. In addition, 2 sows each at 86 days of gestation were inoculated intranasally or IM with MN-Hs virus. ANIMALS: 14 pregnant sows. PROCEDURE: Inoculated sows were allowed to deliver at term, and each litter was examined for clinical abnormality and presence of virus. RESULTS: Two sows infected with MN-Hs virus delivered 14 live and 5 dead pigs, whereas 2 sows infected with MN-HI virus delivered 0 live and 25 dead pigs. Two sows inoculated with a field isolate (MN-W) delivered 10 live and 20 dead pigs. Two control sows had 26 normal fetuses at slaughter at 107 days of gestation. Virus was isolated from 16 (66.7%) of 24 liveborn pigs, 9 (64.3%) of 14 stillborn pigs, and 3 (12.0%) of 25 mummified fetuses of the 6 infected sows. Subsequently, 4 MN-Hs-infected sows delivered 40 live and 11 dead pigs. CONCLUSIONS: Marked difference in the pathogenicity in pregnant sows between porcine reproductive and respiratory syndrome virus strains was documented. The MN-Hs virus is considered to be of low pathogenicity, but the other viruses are highly pathogenic for late-term pregnant sows.

Animals↗

An ELISA for porcine reproductive and respiratory syndrome: production of antigen of high quality.

A reliable method was developed to produce a viral antigen preparation from porcine reproductive and respiratory syndrome virus (PRRSV) infected MARC-145 cells by solubilizing the virus with Triton X-100. This method eliminated problems previously encountered with high background reactions associated with PRRSV antigen or cell control antigen. With this new antigen, an indirect enzyme-linked immunosorbent assay (ELISA) was adapted to detect swine serum anti-body against PRRSV. In the ELISA, non-specific reactions associated with test serum samples have been eliminated by utilizing an effective blocking serum diluent. The ELISA is more sensitive than an indirect immunofluorescent assay (IFA), particularly with late-infection sera, while maintaining a high diagnostic specificity. In a comparison of IFA and ELISA using sera collected from 250 pigs of various ages from 5 herds that had PRRS histories, IFA revealed 178 positive samples and 72 negative samples. All of the IFA-positive sera were proven to be ELISA reactors. However, nearly one-half (34/72) of the IFA-negative samples were also ELISA reactors. The diagnostic specificity and sensitivity of the ELISA were 100% and 96.6% with 257 serum samples collected from known healthy PRRS-negative swine herds and 57 sera collected from infected swine at 6 to 56 days after infection, respectively. The ELISA is technically superior to IFA, time-efficient and cost-effective, and suitable for testing of a large number of samples over a short period of time.

Animals↗

Recurrent reproductive failure associated with porcine reproductive and respiratory syndrome in a swine herd.

An investigation of a case of recurrent reproductive failure in a swine herd was conducted. The subject farm (A) and a neighboring farm (B) had experienced an acute outbreak of porcine reproductive and respiratory syndrome in 1990. During 1992, farm A experienced recurrent episodes of conception failure, stillbirths, preweaning mortality, and sow mortality. Similar clinical findings were not evident on farm B. Evaluation of production records indicated that the problem was occurring in first-parity litters. It was learned that replacement gilts for farm A were being purchased from an outside source, whereas farm B raised its own replacement gilts. Indirect fluorescent antibody testing of gilts from both farms indicated a high prevalence of seronegative gilts on farm A vs a high prevalence of seropositive gilts on farm B. Although there is no correlation between indirect fluorescent antibody titers and immunity, it would appear that exposure to porcine reproductive and respiratory syndrome virus prior to breeding is essential to prevent reproductive disease.

Animals↗

Prevention of the spread of porcine reproductive and respiratory syndrome virus in endemically infected pig herds by nursery depopulation.

An attempt was made to prevent the spread of the virus of porcine reproductive and respiratory syndrome (PRRS) on three Minnesota pig farms that had been experiencing chronic nursing pig problems, including poor growth rates and increased mortality of post weaning pigs. The PRRS virus and different bacterial pathogens were isolated from all three farms during the initial investigation, and all the farms had a high prevalence of PRRS virus-seronegative breeding animals. All the pigs tested within one week after weaning when they were 18 to 22 days old, were seronegative, whereas 80 to 100 per cent of the pigs tested at eight to nine weeks had antibody titres ranging from 1:64 to 1:1024 by an indirect fluorescent antibody method. The seroprevalence among the finishing pigs on the three farms ranged from 25 to 50 per cent. An eradication protocol was established on each farm, involving emptying the nurseries, followed by pumping out the slurry pits and cleaning, washing and disinfecting three times in 14 days. After the nurseries were repopulated there were improvements in nursery mortality and average daily weight gain, and no seropositive animals were detected in the nurseries on any of the farms; the seronegative status was maintained for the six-month testing period.

Animal Husbandry↗

Cloning, expression, and sequence analysis of the ORF4 gene of the porcine reproductive and respiratory syndrome virus MN-1b.

Porcine reproductive and respiratory syndrome virus (PRRSV) MN-1b strain open reading frame 4 (ORF4) has been cloned, sequenced, and expressed in Escherichia coli. The homologies of nucleotide and amino acid sequences between MN-1b (US isolate) and LV (European isolate) are 69% and 64%, respectively. The data also showed that ORF4 of MN-1b is 36 bases shorter than that of LV. Western blot analysis of expressed recombinant ORF4 protein reacted with 65% (26/40) of PRRSV-infected pig sera tested. These results demonstrated that ORF4 of PRRSV may not be a well-conserved region.

Amino Acid Sequence↗

A modified serum neutralization test for the detection of antibody to porcine reproductive and respiratory syndrome virus in swine sera.

Various conditions were evaluated and modified to improve the sensitivity of the serum neutralization (SN) test for detecting antibody in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV). Higher SN titers were consistently obtained by the addition of 20% fresh swine serum to the virus diluent and by the use of a permissive cell clone (MARC-145) derived from the MA-104 cell line. Test sera used to assess the SN test were obtained from 2 groups of 3-week-old pigs infected intranasally with PRRSV (MN-1b). Using the modified method, SN antibody was first detected 9-11 days postinoculation (PI), with a peak evident at 11-21 days PI. The antibody subsequently declined, and a second peak was observed between 41 and 45 days PI. The first antibody peak was not observed and the SN antibody was only detectable between 32 and 41 days PI when the test was done with 20% heated swine serum or without supplemental swine serum. The SN antibody during 2-3 weeks PI was found to be sensitive to 2-mercaptoethanol or anti-swine IgM treatment. The SN antibody titers were high when homologous PRRSV isolate was used in the test but were markedly low for heterologous PRRSV isolates. No difference in antibody titers was observed when homologous and heterologous PRRSV isolates were tested by indirect fluorescent antibody assay. These results indicate that the modified SN method is useful in detecting earlier and higher PRRSV antibody and that it can differentiate among PRRSV isolates.

Animals↗

Enhanced replication of porcine reproductive and respiratory syndrome (PRRS) virus in a homogeneous subpopulation of MA-104 cell line.

Two different cell populations, high- (MARC-145) and low-permissive cell clones (L-1) to porcine reproductive and respiratory syndrome (PRRS) virus, were derived from MA-104 cell line (parent cell: P) by cell cloning. Maximum virus yields in MARC-145, P, and L-1 cell clones were 10(8.5), 10(3.5), and 10(2.5) tissue culture infective dose 50 (TCID50)/0.1 ml, respectively. The MARC-145 cell clone supported replication of all 11 different porcine reproductive and respiratory syndrome virus isolates that were tested. These results indicated that the MARC-145 cells will be useful for PRRS virus replication.

Animals↗

Comparison of porcine alveolar macrophages and CL 2621 for the detection of porcine reproductive and respiratory syndrome (PRRS) virus and anti-PRRS antibody.

The American and European strains of porcine reproductive and respiratory syndrome (PRRS) virus were initially isolated in an established cell line (CL 2621) and porcine alveolar macrophages (PAM), respectively. Subsequent isolation of American strains of this virus in PAM has also been reported. To determine their relative sensitivity for virus isolation, both PAM and CL 2621 cells were inoculated with 98 tissue specimens and 73 serum samples from animals suspected of having PRRS. Four of the 98 tissue samples yielded virus in both cell types, whereas 7 samples were positive only in PAM and 4 samples only in CL 2621. Of the 73 serum samples tested, 18 were positive in PAM of which only 2 were positive in CL 2621. Additionally, 82 isolates obtained initially in CL 2621 were inoculated in PAM cells, and 18 strains isolated originally in PAM were inoculated in CL 2621. Of the 82 CL 2621 isolates, 25 could not be propagated on PAM. Of the 57 that replicated in PAM, as detected by a positive test on indirect fluorescent antibody test, only 28 produced cytopathic effects and 29 did not. Of the 18 PAM isolates, 5 did not grow on CL 2621. Although PAM were relatively more sensitive for virus isolation, their failure to support the growth of certain strains of PRRS virus indicates the existence of variants among PRRS virus strains, and both PAM and CL 2621 should be used for virus isolation from clinical samples. In addition, the sensitivity of these 2 cell types was compared for the detection of fluorescent antibodies to PRRS virus using 179 serum samples from PRRS-infected animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Pathogenesis of porcine reproductive and respiratory syndrome virus infection in mid-gestation sows and fetuses.

Two experiments were undertaken to evaluate whether porcine reproductive and respiratory syndrome (PRRS) virus was able to cross the placenta and infect midgestation fetuses following intranasal inoculation of sows and whether PRRS virus directly infected fetuses following in utero inoculation. In experiment 1, eight sows between 45 and 50 days of gestation were intranasally inoculated with PRRS virus (ATCC VR-2332), and four control sows were inoculated with uninfected cell culture lysate. Virus inoculated sows were viremic on postinoculation (PI) days 1, 3, 5, 7 and 9, shed virus in their feces and nasal secretions, and became leukopenic. Sixty-nine of 71 fetuses from principal sows euthanized on PI day 7, 14 or 21 were alive at necropsy and no virus was isolated from any of the fetuses. Two principal sows that farrowed 65 and 67 days PI delivered 25 live piglets and three stillborn fetuses. The PRRS virus was isolated from two live piglets in one litter. In experiment 2, laparotomies were performed on five sows between 40 and 45 days of gestation and fetuses were inoculated in utero with either PRRS virus alone, PRRS virus plus a swine serum containing PRRS antibodies, or uninfected cell culture lysate. Three sows were euthanized on PI day 4 and two sows on PI day 11. Viral replication occurred in fetuses inoculated with virus alone and was enhanced in fetuses inoculated with virus plus antibody. No virus was isolated from control fetuses.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Characterization of pseudorabies virus antibody responses in young swine after infection and vaccination by using an immunoglobulin M antibody capture enzyme-linked immunosorbent assay.

An immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MACELISA) was developed for the detection of pseudorabies virus (PRV)-specific IgM antibody in swine sera because false-positive reactions frequently occurred when sera from older swine were tested with an indirect IgM enzyme-linked immunosorbent assay. Monoclonal mouse anti-swine IgM was used as the capturing antibody, and rabbit anti-PRV hyperimmune gamma globulin was used as the indicating antibody. Sera from non-PRV-infected, experimentally infected, vaccinated and challenged, passively immune and challenged, and naturally infected swine were evaluated. The PRV MACELISA had a specificity of 95% and was as sensitive and reproducible as previously reported in direct assays. An antibody response was still detectable with the MACELISA 21 days after inoculation. The PRV MACELISA did not detect a consistent antibody response in sera from swine vaccinated with either killed-PRV or modified live-virus vaccines but did detect an antibody response in sera from passively immune pigs after challenge with virulent PRV. These results indicated that the PRV MACELISA may be useful for the rapid serodiagnosis of recent PRV infection in swine.

Animals↗

Isolation of swine infertility and respiratory syndrome virus (isolate ATCC VR-2332) in North America and experimental reproduction of the disease in gnotobiotic pigs.

A recent epizootic of swine infertility and respiratory syndrome (SIRS) in a Minnesota swine herd was investigated. Examination of a sow, neonatal piglets, and stillborn fetuses obtained during the epizootic from the affected herd revealed interstitial pneumonitis, lymphomononuclear encephalitis, and lymphomononuclear myocarditis in the piglets and focal vasculitis in the brain of the sow. Fetuses did not have microscopic lesions. No cause for the infertility and respiratory syndrome was determined. Therefore, attempts were made to experimentally reproduce the disease. Eleven 3-day-old gnotobiotic piglets exposed intranasally to tissue homogenates of piglets from the epizootic became inappetent and febrile by 2-4 days postexposure and had interstitial pneumonitis and encephalitis similar to that seen in the field outbreak. After 2 blind passages in gnotobiotic piglets, tissue homogenates were cultured on continuous cell line CL2621, and a cytopathic virus (ATCC VR-2332), provisionally named SIRS virus, was isolated. Gnotobiotic piglets exposed intranasally to the SIRS virus developed clinical signs and microscopic lesions that were the same as those in piglets exposed to the tissue homogenates, and the virus was reisolated from their lungs. This is the first isolate of SIRS virus in the United States that fulfills Koch's postulates in producing the respiratory form of the disease in gnotobiotic piglets and the first report of isolation and propagation of the virus on a continuous cell line (CL2621). The virus is designated as American Type Culture Collection VR-2332.

Animals↗

Isolation of a cytopathic virus from weak pigs on farms with a history of swine infertility and respiratory syndrome.

Severe clinical signs of swine infertility and respiratory syndrome (SIRS) of unknown cause were observed in several Minnesota swine farms between November 1990 and March 1991. Forty-five lung samples of weak pigs were collected from 13 swine farms, and virus isolation was attempted using swine alveolar macrophage (SAM) cultures. A cytopathic virus was isolated from 19 lung samples collected from 6 different farms. Four pregnant sows were infected intranasally with a tissue suspension from which virus was isolated, and 4 6-week-old pigs and 2 contact pigs were infected intranasally with 1 of the isolates. The 4 sows farrowed 12 stillborn and 32 normal pigs. Virus was recovered from 10 of 19 pigs examined. Infected 6-week-old pigs were clinically normal except for slightly elevated rectal temperatures and mild respiratory signs. No or mild interstitial pneumonic lesions were observed in inoculated pigs, but the lesion was obvious in the 2 contact pigs. Seroconversion was observed in sows and pigs as measured by indirect fluorescent antibody (IFA). Serologic identification of the isolates was carried out by IFA using reference serum prepared from an experimentally infected sow. A cytoplasmic fluorescence was observed on the SAM monolayers infected with each of the 19 different isolates. Fluorescence was also observed when the monolayers were tested with SIRS virus ATCC VR-2332-infected sow sera. Replication of the isolates was not affected in the medium containing 5-iodo-2'-deoxyuridine but was inhibited by treatment with ether. The isolates were relatively stable at 56 C and did not agglutinate with various erythrocytes tested.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

An indirect fluorescent antibody test for the detection of antibody to swine infertility and respiratory syndrome virus in swine sera.

An indirect fluorescent antibody (IFA) test was developed and standardized to detect and quantitate antibody for swine infertility and respiratory syndrome (SIRS) virus in swine sera. Test results were evaluated using sera of pigs infected both experimentally and naturally with SIRS virus. The IFA test used swine alveolar macrophage (SAM) monolayers prepared in 96-well microplates and infected with SIRS virus. The monolayers were incubated with test sera, washed, and stained with fluorescein isothiocyanate-labeled rabbit anti-swine IgG. After another wash step, the monolayers were examined under a fluorescent microscope. A noninfected SAM control well was included for each sample. The antibody titers for each serum sample were recorded as the highest serum dilutions with specific cytoplasmic fluorescence but no fluorescence in the control wells. To evaluate the test, sera of 4 6-week-old pigs that had been infected with SIRS virus, 2 contact pigs, and 13 experimentally infected sows were used. In the experimentally infected pigs, antibody was first detected at 7 days postexposure (PE) and peaked (1:256-1,024) between 11 and 21 days PE. All 13 sow sera were negative at time of infection but were positive (1:64- greater than or equal to 1:1,024) at 14-26 days PE. Seven hundred twenty sera collected from 25 different swine farms with or without a history of SIRS were also tested. Of 344 sera from 15 swine farms with a clinical history of SIRS, 257 (74.7%) sera had IFA titers greater than or equal to 1:4, whereas 371 (98.7%) of 376 sera from herds with no history of SIRS were negative.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

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.

Animals↗