Immunofluorescent Studies of Bovine Para-Influenza 3 Virus II. Experimentally Infected Calves

Fluorescent antibody (FA) studies of tissues from three colostrum deprived calves inoculated intranasally with the SF-4 strain of bovine para-influenza 3 (PI-3) virus indicated that these calves developed a mild upper respiratory infection but infected cells were not identified in the lower respiratory tract. Three other calves inoculated intranasally and intratracheally with PI-3 virus developed more severe clinical signs of infection and virus was identified, by FA techniques, in the upper and lower respiratory tract of all three calves and in the spleen of one calf. PI-3 virus was detected in smears of nasal epithelium from five of six calves at some time during the observation period.     

Serological studies of parainfluenza type 3 virus, bovine adenovirus type 3 and bovine respiratory syncytial virus infection in beef calves

Six serum samples were taken at monthly intervals from birth to weaning from each of 41 newborn calves in the autumn and spring calf crops of a beef cow--calf herd. The serum hemagglutination-inhibition (HI) antibody titres to parainfluenza type 3 virus (PIV-3), virus-neutralization (VN) antibody titres to bovine adenovirus type 3 (BAV-3) and bovine respiratory syncytial virus (BRSV) were determined using microtitration techniques. There was serological evidence of a significantly higher incidence of infection with BAV-3 in the fall calves than in the spring calves. Serological responses to BAV-3 were not detected in calves with VN titres of greater than 1/256. Serological evidence of subclinical infection with PIV-3 occurred mainly in late February or early March during a period of marked environmental temperature fluctuations. Serological evidence of a high incidence of infection with BRSV was obtained for both the fall and spring calf crops. Serum antibody appeared to be unable to prevent infection with BRSV. An association between infection with BRSV and clinical pneumonia was found in 3 out of 9 calves. BAV-3 infection was related to pneumonia in only 1 instance; however, there was simultaneous evidence of BRSV infection in this calf. PIV-3 infection was found to be related to pneumonia in only 1 instance. There was serological evidence of infection with BAV-3 in association with the occurrence of diarrhea in 3 calves.     

Comparison of a newly developed enzyme-linked immunosorbent assay with complement fixation and neutralisation tests for serology of bovine respiratory syncytial virus infections

An indirect double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the detection and titration of serum antibodies to bovine respiratory syncytial virus (BRSV). The ELISA was compared with a complement fixation (CF) test and a test for virus neutralising antibody in serum (virus neutralisation [VN] test). Testing sera collected in dairy herds revealed the closest correlation between the results of the ELISA and the CF test with respect to BRSV antibody titres. The VN test detected BRSV antibodies in a higher percentage of acute phase sera compared to the other two tests in field samples and in early bleedings of experimentally infected calves. However, the VN test was less effective in making a diagnosis of BRSV infections on the basis of a significant titre increase in paired sera. For this purpose the ELISA was found to be the most sensitive test.     

Efficacy of viral components of a nonabortigenic combination vaccine for prevention of respiratory and reproductive system diseases in cattle

Efficacy and safety of components of an IM-administered vaccine for prevention of infectious bovine rhinotracheitis virus (IBRV), parainfluenza type-3 (PI-3) virus, bovine viral diarrhea virus (BVDV), and respiratory syncytial virus (RSV) infections and campylobacteriosis and leptospirosis were evaluated in cattle, including calves and pregnant cows. Challenge of immunity tests were conducted in calves for IBRV, PI-3 virus, or BVDV vaccinal components. All inoculated calves developed serum-neutralizing antibodies and had substantially greater protection (as measured by clinical rating systems) than did controls after challenge exposure to virulent strains of IBRV, PI-3 virus, BVDV, or RSV. In in utero tests, IBRV or bovine RSV vaccinal strains were inoculated into fetuses of pregnant cows. Histologic changes or abortions did not occur after fetal inoculation of the RSV vaccinal strain, and 10 of 14 fetuses responded serologically. Of 9 fetuses, one responded serologically to the IBRV vaccinal strain after in utero inoculation and was aborted 3 weeks later. In an immunologic interference test, 10 calves vaccinated with 2 doses of the multivalent vaccine, containing the 4 viral components and a Campylobacter-Leptospira bacterin, developed serum-neutralizing antibodies to IBRV, PI-3 virus, BVDV, and RSV without evidence of serologic interference. Under field conditions, 10,771 cattle, including 4,543 pregnant cows, were vaccinated. Vaccine-related abortions did not occur.     

Antibody response of calves to immunoaffinity-purified bovine respiratory syncytial virus VP70 after vaccination and challenge exposure.

Immunoaffinity-purified bovine respiratory syncytial virus (BRSV) fusion (F) protein elicited anti-BRSV-specific antibody responses in BRSV-seronegative calves. After primary vaccination, all calves seroconverted to BRSV as determined by the virus neutralization (VN) test and developed anti-F protein antibodies detectable by protein immunoblot analyses. Subsequent vaccinations induced greater than twofold increase in VN titer in 3 of 9 (33%) calves, and 1 calf became VN-negative, but still had nonneutralizing antibody detectable by protein immunoblot analysis. This calf remained seronegative after challenge exposure. Two groups of calves were vaccinated IM with immunoaffinity-purified BRSV F protein. Each dose was 2 ml containing 20 micrograms of purified F protein. Freund's adjuvants were used for all vaccinations, with Freund's complete adjuvant used for the primary vaccination and Freund's incomplete adjuvant for subsequent vaccinations. The vaccine was administered to both groups at weeks 0 and 3; the first group received a third vaccination at weeks 21. Group-1 and -2 vaccinated calves and non-vaccinated contact controls were intranasally aerosol challenge-exposed with low cell culture-passage BRSV on weeks 22 and 9, respectively. Eight of 9 vaccinated calves did not develop a humoral anamnestic response following challenge exposure, as demonstrated by VN test and protein immunoblot analyses. Calf 14 from group 1 which had a 1:2 VN antibody titer prior to vaccination, was the only calf that developed an anamnestic response.(ABSTRACT TRUNCATED AT 250 WORDS)     

Clinical and immunologic responses of calves with colostrally acquired maternal antibody against parainfluenza-3 virus to homologous viral infection.

Exposure of colostrum-deprived calves and calves with colostrally acquired maternal antibody to aerosols of parainfluenza-3 (PI-3) virus resulted in signs of infection, leukopenia, and shedding of virus from the nasal passages. However, infection was not as severe in calves with colostrally acquired maternal antibody as it was in colostrum-deprived calves which did not have antibody to PI-3 virus before they were exposed. All calves responded immunologically to PI-3 virus, as indicated by resistance to challenge exposure and subsequent development of virus-neutralizing antibody. However, levels of serum and nasal secretion (NS) antibody at 30 days after viral exposure were lower in calves with colostrally acquired maternal antibody than in colostrum-deprived calves, and a serum antibody response in the former was primarily indicated by an anamnestic response after challenge exposure. After calves were challenge exposed to PI-3 virus, serum and NS antibodies were increased in all calves, but antibody titers were generally lower for calves that had colostrally acquired maternal antibody before their exposure than for those that acquired antibody only after PI-3 viral infection.     

Investigation of causative agents of bovine respiratory tract disease in a beef cow-calf herd with an early weaning program.

Serum samples were collected from early weaned fall calves shortly after the onset of respiratory tract disease. Antibody titers to infectious bovine rhinotracheitis (IBR) virus, parainfluenza type 3 (PI-3) virus, bovine viral diarrhea (BVD) virus, bovine adenovirus type 3 (BAV-3), and bovine respiratory syncytial virus (BRSV) were determined on paired (acute and convalescent) serums. Seroconversion rate (a fourfold or greater rise in antibody titer) for IBR virus was 4.3%, PI-3 virus--16.3%, BVD virus--9.6%, and BAV-3--2.2%. Seroconversion for BRSV was 45.4%. An increased rate of seroconversion for IBR, PI-3, and BVD viruses and BAV-3 was observed in the presence of BRSV seroconversion. These results suggest that BRSV may facilitate infection by other viruses. Results of virus isolation procedures from these calves were negative.     

Epidemiologic studies of the detection of viral infection agents in calf losses in the Weser-Ems region

In 961 calves up to an age of 6 months which were sent to the animal health center in Oldenburg between March 1987 and March 1990 for necropsy the results of determination of different viruses were calculated: BVD-, rota-, corona-, parainfluenza-3- (PI-3)-, bovine herpes-1 (BHV-1)- and bovine respiratory syncytial virus (BRSV). In 122 and 104 randomly collected health calves of 22 farms antibodies against BRSV and bovine adeno virus-types 5, 7 and 8 were determined. 50.1% of the necropsied calves were one and two weeks old. In this group in 40.2% rotavirus and in 19.0% coronavirus could be isolated. All over the calves the frequencies of isolated viruses were 13.3% for BVDV, 4.6% for BRSV, 3.2% for BHV-1, and 2.1% for PI-3. The percentages of positive findings for rota- and coronavirus increased up to 7 days after birth, and thereafter both decreased. The frequencies of BVDV and BRSV were higher in older groups. The frequency of PI-3 was low and remained constantly. Infections with rota-, corona- and with both viruses were accompanied by BVDV in 11.3, 5.3 and 14.3%, respectively. Against bovine adenoviruses and BRSV in the first 8 weeks and after 14 weeks of life in more than 70% of the calves antibodies were detected.     

Bovine viral diarrhea viral infections in feeder calves with respiratory disease: interactions with Pasteurella spp., parainfluenza-3 virus, and bovine respiratory syncytial virus.

The prevalence of bovine viral diarrhea virus (BVDV) infections was determined in a group of stocker calves suffering from acute respiratory disease. The calves were assembled after purchase from Tennessee auctions and transported to western Texas. Of the 120 calves, 105 (87.5%) were treated for respiratory disease. Sixteen calves died during the study (13.3%). The calves received a modified live virus BHV-1 vaccine on day 0 of the study. During the study, approximately 5 wk in duration, sera from the cattle, collected at weekly intervals, were tested for BVDV by cell culture. Sera were also tested for neutralizing antibodies to BVDV types 1 and 2, bovine herpesvirus-1 (BHV-1), parainfluenza-3 virus (PI-3V), and bovine respiratory syncytial virus (BRSV). The lungs from the 16 calves that died during the study were collected and examined by histopathology, and lung homogenates were inoculated onto cell cultures for virus isolation. There were no calves persistently infected with BVDV detected in the study, as no animals were viremic on day 0, nor were any animals viremic at the 2 subsequent serum collections. There were, however, 4 animals with BVDV type 1 noncytopathic (NCP) strains in the sera from subsequent collections. Viruses were isolated from 9 lungs: 7 with PI-3V, 1 with NCP BVDV type 1, and 1 with both BVHV-1 and BVDV. The predominant bacterial species isolated from these lungs was Pasteurella haemolytica serotype 1. There was serologic evidence of infection with BVDV types 1 and 2, PI-3V, and BRSV, as noted by seroconversion (> or = 4-fold rise in antibody titer) in day 0 to day 34 samples collected from the 104 survivors: 40/104 (38.5%) to BVDV type 1; 29/104 (27.9%) to BVDV type 2; 71/104 (68.3%) to PI-3V; and 81/104 (77.9%) to BRSV. In several cases, the BVDV type 2 antibody titers may have been due to crossreacting BVDV type 1 antibodies; however, in 7 calves the BVDV type 2 antibodies were higher, indicating BVDV type 2 infection. At the outset of the study, the 120 calves were at risk (susceptible to viral infections) on day 0 because they were seronegative to the viruses: 98/120 (81.7%), < 1:4 to BVDV type 1; 104/120 (86.7%) < 1:4 to BVDV type 2; 86/120 (71.7%) < 1:4 to PI-3V; 87/120 (72.5%) < 1:4 to BRSV; and 111/120 (92.5%) < 1:10 to BHV-1. The results of this study indicate that BVDV types 1 and 2 are involved in acute respiratory disease of calves with pneumonic pasteurellosis. The BVDV may be detected by virus isolation from sera and/or lung tissues and by serology. The BVDV infections occurred in conjunction with infections by other viruses associated with respiratory disease, namely, PI-3V and BRSV. These other viruses may occur singly or in combination with each other. Also, the study indicates that purchased calves may be highly susceptible, after weaning, to infections by BHV-1, BVDV types 1 and 2, PI-3V, and BRSV early in the marketing channel.     

Field application of the combination of neutralizing test and virus isolation, so-called spot test, to detect herds including cattle persistently infected with bovine viral diarrhea virus

To detect herds including cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV), application of the combination of neutralizing antibody detection and virus isolation, so-called spot test, were performed on sera of 3 calves selected from each of 26 farms. Nine farms were judged as positive because 64 or more antibody titers were detected from 2 or more calves or BVDV was isolated from one or more calves. PI cattle were detected from 8 of the 9 farms. The positive judgment on one farm was obtained only when the indicator virus used on the neutralizing test was genotypically identical with the isolate from the farm. These results suggest that the spot test can be effective in detecting herds with PI cattle and that the accuracy may be influenced by the genotypes of the indicator viruses.     

Protective immunity against Newcastle disease: the role of cell-mediated immunity

The role of cell-mediated immunity (CMI) in protection of birds from Newcastle disease was investigated by two different strategies in which only Newcastle disease virus (NDV)-specific CMI was conveyed without neutralizing antibodies. In the first strategy, selected 3-wk-old specific-pathogen-free (SPF) birds were vaccinated with either live NDV (LNDV), ultraviolet-inactivated NDV (UVNDV), sodium dodecyl sulfate-treated NDV (SDSNDV), or phosphate-buffered saline (PBS) (negative control) by the subcutaneous route. Birds were booster vaccinated 2 wk later and challenged with the velogenic Texas GB strain of NDV 1 wk after booster. All vaccinated birds had specific CMI responses to NDV as measured by a blastogenesis microassay. NDV neutralizing (VN) and hemagglutination inhibition (HI) antibody responses were detected in birds vaccinated with LNDV and UVNDV. However, birds vaccinated with SDSNDV developed antibodies that were detected by western blot analysis but not by the VN or HI test. Protection from challenge was observed only in those birds that had VN or HI antibody response. That is, birds with demonstrable CMI and VN or HI antibody response were protected, whereas birds with demonstrable CMI but no VN or HI antibody response were not protected. In the second strategy, birds from SPF embryos were treated in ovo with cyclophosphamide (CY) to deplete immune cells. The birds were monitored and, at 2 wk of age, were selected for the presence of T-cell activity and the absence of B-cell activity. Birds that had a significant T-cell response, but not a B-cell response, were vaccinated with either LNDV, UVNDV, or PBS at 3 wk of age along with the corresponding CY-untreated control birds. The birds were booster vaccinated at 5 wk of age and were challenged with Texas GB strain of NDV at 6 wk of age. All birds vaccinated with LNDV or UVNDV had a specific CMI response to NDV, VN or HI NDV antibodies were detected in all CY-nontreated vaccinated birds and some of the CY-treated vaccinated birds that were found to have regenerated their B-cell function at 1 wk postbooster. The challenge results clearly revealed that CY-treated birds that had NDV-specific CMI and VN or HI antibody responses to LNDV or UVNDV were protected, as were the CY-nontreated vaccinated birds. However, birds that had NDV-specific CMI response but did not have VN or HI antibodies were not protected from challenge. The results from both strategies indicate that specific CMI to NDV by itself is not protective against virulent NDV challenge. The presence of VN or HI antibodies is necessary in providing protection from Newcastle disease.     

Prevalence of influenza viruses A-H1N1 and A-H3N2 in swine in The Netherlands

In the period December 1979-May 1980 a respiratory disease spread rapidly through pig herds in The Netherlands. Surveillance of 12 pig farms resulted in isolation of 22 influenza A-Swine-H1N1 (Hsw1N1) strains from 9 pig herds. The morbidity rate was high but the mortality rate was nil. Retardation in growth was observed. Sera collected from affected pig herds showed a fourfold increase in haemagglutination inhibition (HI) titre against A-Swine-H1N1 virus. Sera collected on five farms showed a geometric mean HI titre against the A-H3N2 virus above 100. A significant HI titre increase against this virus was found in sera collected on three farms. These findings indicated a recent infection by this virus. A-H3N2 virus was not isolated. The Dutch Swine-1980 isolates showed in the cross-HI test a distant antigenic relationship with the classical A/Swine/Iowa/30 (H1N1) virus and one-sided close antigenic relationship with A/New Jersey/76 (H1N1) virus.HI antibody to A/Swine/Nederland/80 (H1N1) virus was found in 4, 0, and 44%, to A/New Jersey/76 (H1N1) virus in 0.5, 0.4, and 42%, and to A/Swine/Iowa/30 (H1N1) virus in 0.5, 1, and 30% of pig sera collected in 1976, 1977, and 1980, respectively. HI antibody to A/Hong Kong/68 (H3N2) virus was detected in 36, 56, and 68%, and to A/Victoria/75 (H3N2) virus in 38, 73, and 68% of these sera, respectively.The results of this study indicate that pigs in The Netherlands, like those in North America, Southeast Asia, Japan, and Western Europe harbour A-Swine-H1N1 and A-H3N2 influenza viruses and are thus potential reservoirs for future human pandemics.     

Antibody response of calves after intranasal inoculation with parainfluenza-3 virus and resistance of inoculated calves to experimental homologous viral infection

The immunologic response of colostrum-deprived calves to parainfluenza-3 (PI-3) virus given by intranasal inoculation was studied. Inoculation of calves with 3.2 x 10(6) median cell culture infective doses (CCID50) of either virulent (SF-4) virus or a modified strain of PI-3 virus, or with 2.0 x 10(8) CCID50 of SF-4 virus, stimulated development of both serum antibody and nasal secretion (NS) antibody. However, NA antibody decreased in all calves between the 16th and 42nd postinoculation days and was present only at low or moderately low concentrations on the 126th day, when the immunity of the calves was challenged. Generally, calves that were inoculated with 3.2 x 10(6) CCID50 of SF-4 virus developed slightly higher concentrations of serum and NS antibodies than did calves inoculated with modified virus. Calves that were inoculated with 2.0 x 10(8) or 3.2 x 10(6) CCID 50 of SF-4 virus developed comparable concentrations of serum antibody, but large doses of SF-4 virus were less effective than smaller doses of the same virus in stimulating the development of NS antibody. Reinoculation of 3 calves with modified PI-3 virus resulted in a demonstrable increase in serum antibody in 2 calves and an increase and subsequent decrease in NS antibody in all calves. Challenge exposure of inoculated calves to aerosols of SF-4 virus failed to cause clinical signs of disease, and the challenge virus was not isolated from the nasal passages.     

Correlation between maternal serum antibodies and protection against bovine rotavirus diarrhea in calves

The correlation between maternal serum antibodies in beef calves at 2 days old and protection against diarrhea induced by natural bovine rotavirus (BRV) infection was examined. Virus neutralizing (VN) antibody titers against BRV in sera from calves that developed diarrhea by BRV infection within 14 days of age (BRV-diarrheal calves) were significantly lower than those from calves that had no diarrhea. In the BRV-diarrheal calves, a positive correlation was found between the VN antibody titers and age of the onset of diarrhea. There were negative correlations between the VN antibody titers and duration of the diarrhea, VN antibody titers and cumulative diarrhea scores, and the VN antibody titers and duration of virus shedding. These results suggest that the VN antibody titers against BRV in newborn calf serum could be an indicator of protection against BRV-induced diarrhea.     

The prevalence of bovine viral diarrhea virus infection in a population of feedlot calves in western Canada.

The prevalence of bovine viral diarrhea virus (BVDV) infection was examined in a population of 5129 recently weaned steer calves entering a large feedlot in central Saskatchewan from September to December 1991. Serum samples were collected within 24 h of arrival at the feedlot from every fifth calf processed and again 96 d postarrival. A microtiter virus isolation test was used to determine the prevalence of calves viremic with BVDV on entry to the feedlot. An enzyme-linked immunosorbent assay (ELISA) which detects antibody against glycoprotein 53 of the BVDV was used on paired sera to determine the seroconversion risk during the first 96 d in the feedlot. A virus neutralization (VN) test for BVDV was conducted on a sub-sample of paired sera to measure agreement in determination of seroconversion risk with the ELISA. A polymerase chain reaction (PCR) test which detects BVDV was used to determine if cattle were acutely viremic when treated for disease. The estimated prevalence of persistently infected calves in this population was < 0.1%. The seroconversion risk for BVDV was 27% (236/864) according to the ELISA and it varied from 0 to 63% among the 20 pens sampled. According to the VN test, the seroconversion risk for BVDV was 40% (132/327) and it varied from 0 to 100% among the 11 pens tested. The agreement between the ELISA and VN tests in seroconversion risk to BVDV was very poor (kappa = 0.15 +/- 0.039 SE). The prevalence of acute viremia in calves treated at the feedlot hospital was low at 4% (6/149).(ABSTRACT TRUNCATED AT 250 WORDS)     

Lack of antigenic relationships between avian pneumovirus and four avian paramyxoviruses

Avian paramyxoviruses (PMVs) and avian pneumovirus (APV) belong to the family Paramyxoviridae. Antigenic relationships between PMVs were shown previously, hence, this study was designed to investigate possible antigenic relationships between APV and four avian PMVs (PMV-1, PMV-2, PMV-3, and PMV-7). Enzyme-linked immunosorbent assay (ELISA), hemagglutination inhibition (HI) test, and virus neutralization (VN) test in chicken embryos and in Vero cells were used. The HI test was performed with the PMVs as antigens against the APV and PMVs antisera. The ELISA and VN test in chicken embryos were performed with PMVs and APV antigens and antisera. The VN test in vero cells was performed with the APV as an antigen against the PMV antisera. All the viruses were isolated in the United States or Canada. No antigenic relationships between APV and the PMVs were detected by the described tests.     

Prevalence of Pasteurella multocida and other respiratory pathogens in the nasal tract of Scottish calves

The prevalence of Pasteurella multocida, a cause of bovine respiratory disease, was studied in a random sample of beef suckler and dairy farms throughout Scotland, by means of a cross-sectional survey. A total of 637 calves from 68 farms from six geographical regions of Scotland were sampled between February and June 2008. Deep nasal swabs were taken, and samples that were culture-positive for P multocida were confirmed by PCR. Prevalence of P multocida was 17 per cent (105 of 616 calves); 47 per cent of farms had at least one positive animal. A higher prevalence was detected in dairy calves than beef calves (P=0.04). It was found that P multocida was associated with Mycoplasma-like organisms (P=0.06) and bovine parainfluenza type 3 virus (BPI-3) (P=0.04), detected by culture and quantitative PCR of nasal swabs, respectively. Detection of P multocida was not associated with bovine respiratory syncytial virus (BRSV), bovine herpesvirus type 1 (BoHV-1) or bovine viral diarrhoea virus (BVDV). Mycoplasma-like organisms, BPI-3, BRSV, BoHV-1 and BVDV were detected in 58, 17, four, 0 and eight calves, on 25, five, two, 0 and five of the 68 farms, respectively.     

Functional analysis of antibody responses of feedlot cattle to bovine respiratory syncytial virus following vaccination with mixed vaccines.

The antibody response of cattle to bovine respiratory syncytial virus (BRSV) immunization was investigated using 4 different commercially available mixed vaccines. Forty, 5-6 month old, beef calves, randomly assigned to groups of 10, were vaccinated on day 0 and 21 with 1 of 3 inactivated vaccines, (3 groups), or a modified live virus (MLV) vaccine. BRSV-specific antibody responses were measured prior to vaccination and on day 35 by using an enzyme linked immunosorbent assay (ELISA), virus neutralization assay (VN), a fusion inhibition assay (FI); and responses were also measured for their ability to facilitate antibody dependent, complement mediated cytotoxicity (ADCMC) of BRSV infected cells. Sera from day 35 were, in addition, analyzed by use of an IgG1, IgG2 isotype specific ELISA. All vaccines induced significant increases in BRSV specific IgG antibody as measured by ELISA, but only one inactivated and the MLV vaccine induced significant increases in VN titers. Fusion inhibiting antibody titers were low or undetected in calves vaccinated with the inactivated vaccines. Vaccination with modified live virus induced significantly higher titers of fusion inhibiting antibodies, which are considered to be most highly correlated with protection. The VN to ELISA and FI to ELISA ratio of the calves that received MLV vaccine were significantly greater than the calves receiving the 3 inactivated vaccines. Vaccination with MLV induced the highest IgG2/IgG1 ratio. This difference was small, and only significant relative to 2 of the inactivated vaccine groups, which were not significantly different from each other. The higher proportion of IgG2 isotype in the MLV sera was not associated with lower ADCMC, a function not attributed to this isotype. The VN and FI titers, but not the ELISA value of the sera, were most predictive of ADCMC. The inactivation processes apparently alter epitopes and affect the induction of functional antibodies.     

Evaluation of enzyme-linked immunosorbent assay (ELISA) and immunofluorescent antibody (IFA) test for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibody in pigs from conventional farms

To evaluate the immunofluorescent antibody (IFA) test and enzyme-linked immunosorbent assay (ELISA) for detecting the porcine reproductive and respiratory syndrome virus (PRRSV) antibody, conventional pigs in PRRSV-positive and -negative commercial farms were examined. Antibody development patterns in ELISA and IFA tests were compared in 3 week old piglets experimentally infected with the PRRSV. The virus was detected from 2 days post infection (PI) and then the antibody titers and S/P ratios rose by both methods. A total of 208 serum samples were collected from 4 PRRSV-negative farms and 210 samples from PRRSV-positive farms, and were tested for the PRRSV antibody by IFA and ELISA. The titer of 64 should be set as the cut-off point in IFA for field sera. Similarly, the cut-off S/P ratio should be set at 0.4 in ELISA. A high degree of correlation was observed between antibody titers by the two methods in these 418 samples, with a correlation coefficient of 0.84. The coincidence rate between the two tests was 84.7% (354/418). In non-coincident cases, ELISA was able to detect the antibody with a low titer in the serum samples which were negative in IFA but from PRRSV positive farms. ELISA was more sensitive than IFA to detect PRRSV infected animals or farms.     

The development of antibodies in rabbits and cattle infected experimentally with an african strain of malignant catarrhal fever virus

An indirect immunofluorescence (IIF) test for antibody to wildebeest-derived strains of malignant catarrhal fever virus (MCFV) was developed using infected bovine embryonic kidney cells as antigen. This IIF technique and a microtitre method for the assay of virus-neutralizing (VN) antibody were then applied to sequential serum samples from rabbits and calves infected experimentally with a virulent strain of MCFV, which caused lethal disease.Antibody of the IIF type appeared in both species about 6–7 days prior to the onset of pyrexia and increased continuously until death. Neutralizing antibody was detected shortly before the end of the incubation period in the majority of rabbits but was not produced in calves which were killed up to 3 days after reaction. The presence of either antibody did not affect the fatal outcome in rabbits.