Residual viruses in pork products.

Partly cooked canned hams and dried pepperoni and salami sausages were prepared from the carcasses of pigs infected with African swine fever virus and pigs infected with hog cholera virus. Virus was not recovered from the partly cooked canned hams; however, virus was recovered in the hams before heating in both instances. Both African swine fever virus and hog cholera virus were recovered from the dried salami and pepperoni sausages, but not after the required curing period.     

Susceptibility of mink to certain viral animal diseases foreign to the United States.

Mink (Mustela vison) were inoculated with viruses: African horse sickness (AHS), African swine fever (ASF), bovine herpes virus II (BHV2), foot-and-mouth disease (FMD), goat pox (GP), hog cholera (HC), peste des petits ruminants (PPR), rinderpest (RP), swine vesicular disease (SVD), vesicular exanthema of swine (VES) and vesicular stomatitis (VS). Their susceptibility was measured by development of clinical signs, virus isolation and detection of precipitin and/or virus neutralizing antibodies. SVD virus produced a lesion in one mink. Virus was isolated from mink inoculated with SVD, FMD and BHV2. Neutralizing and/or precipitin antibodies were detected in mink inoculated with ASF, FMD, GP, RP, SVD and VS viruses. Mink were not susceptible to AHS, HC, PPR and VES viruses.     

Live attenuated pseudorabies virus expressing envelope glycoprotein E1 of hog cholera virus protects swine against both pseudorabies and hog cholera

To investigate whether live attenuated pseudorabies virus (PRV) can be used as a vaccine vector, PRV recombinants that expressed envelope glycoprotein E1 of hog cholera virus (HCV) were generated. Pigs inoculated with these recombinants developed high levels of neutralizing antibodies against PRV and HCV and were protected against both pseudorabies and hog cholera (classical swine fever).     

Inactivation of viral agents in bovine serum by gamma irradiation

Cell culture origin or suckling mouse brain origin viruses of Akabane disease, Aino, bovine ephemeral fever, swine vesicular disease, hog cholera, bluetongue, and minute virus of mice were each suspended in bovine serum. Aliquots (1 mL) were exposed to various doses of gamma radiation from a 60Co source while at -68 degrees C. Aliquots (100-mL) of serum from a steer experimentally infected with foot-and-mouth disease virus were similarly irradiated. The samples were assayed for infectivity in cell culture systems before and after irradiation, and the data points were analyzed by linear regression. The irradiation doses (in megarads) necessary to inactivate one log10 of viral infectivity (D10) was calculated for each virus. D10 is otherwise known as the slope of the regression line. The r2 value, a measure of association with 1.0 = perfect fit, was also calculated for each regression line. The values (D10, r2) for each virus were as follows: Akabane, 0.25, 0.998; Aino, 0.35, 0.997; bovine ephemeral fever, 0.29, 0.961; swine vesicular disease, 0.50, 0.969; foot-and-mouth disease, 0.53, 0.978; hog cholera, 0.55, 0.974; bluetongue, 0.83, 0.958; and minute virus of mice, 1.07, 0.935.     

Identification of Hog Cholera Viral Antigen by Immunofluorescence. Application as a Diagnostic and Assay Method

In the absence of detectable cytologic changes in hog cholera virus-infected tissue culture cells, hog cholera viral antigen was readily detected by immunofluorescence. The ability to detect hog cholera viral antigen by this method allowed for determination of infectivity titers and also for titration of homologous antibody. Immunofluorescence made possible the identification, in tissue culture, of hog cholera virus from blood, serum, and spleen extracts of experimentally infected swine. Further applications of this method and its limitations are being investigated.     

Vaccination of Pigs Against Hog Cholera (Classical Swine Fever) With a Detergent Split Vaccine

Four pigs were inoculated subcutaneously with a detergent (triton X 100) split hog cholera virus in Freund’s incomplete adjuvant. Four other pigs were in the same way inoculated with a detergent split bovine viral diarrhoea virus, also in Freund’s incomplete adjuvant. In the experiment were used 3 control pigs. The vaccinations were repeated after 3 weeks. All pigs were challenged with highly virulent hog cholera virus (Tübingen) 12 weeks after primary inoculations. Signs of hog cholera were only noted in the control pigs. This introductory experiment was succeeded by a larger experiment with subcutaneous inoculations of: 10 pigs with detergent split hog cholera virus in Freund’s incomplete adjuvant, 10 pigs with detergent split hog cholera virus in a saponin (Quil A) solution, 10 pigs with detergent split bovine viral diarrhoea virus in Freund’s incomplete adjuvant, 10 pigs with detergent split bovine viral diarrhoea virus in the Quil A solution plus 5 control pigs. The vaccinations were repeated after 3 weeks, and finally all pigs were challenged 9 weeks later with the highly virulent hog cholera virus strain. With the exception of 1 animal which died accidentally, all animals survived in the groups inoculated with the Quil A vaccines and in the group inoculated with the detergent split hog cholera virus/oil adjuvant vaccine. In the group inoculated with the detergent split bovine viral diarrhoea virus/oil adjuvant vaccine, some of the pigs died of hog cholera.     

Glutaraldehyde inactivation of exotic animal viruses in swine heart tissue.

Glutaraldehyde, 0.2%, in a 1:100 (wt/vol) ratio, inactivated four animal viruses (foot-and-mouth disease, swine vesicular disease, African swine fever, hog cholera) in swine heart tissues during 11-day exposures at 22 to 26 degrees C.     

Differences in pathogenicity and antigenicity among hog cholera virus strains.

Using antiserum against a particular strain of bovine viral diarrhea virus, the strains of hog cholera virus were divided into two groups, H and B, on the basis of the difference in the degree of neutralization. Group H consisted of strains reacting poorly in neutralization, and group B Consisted of strains reacting well with bovine viral diarrhea antiserum. Most of the strains of group H induced a typical clinical form of hog cholera in experimentally infected pigs. Inoculation of pigs with a strain of group B, however, resulted in a chronic type of illness. When immunized with bovine viral diarrhea virus, pigs succumbed to challenge with group H virus after showing clinical signs of hog cholera, but survived challenge with group B virus without manifesting any clinical sign.     

Demonstration of an Antigenic Relationship Between Hog Cholera and Bovine Viral Diarrhea Viruses by Immunofluorescence

Specific staining of antigen within bovine embryo kidney tissue culture cells, infected with either Oregon C24V or NADL-MD bovine viral diarrhea virus, was accomplished using fluorescein-conjugated swine anti-hog cholera or bovine antiviral diarrhea globulin. Also specific staining of antigen within pig kidney tissue culture cells, infected with hog cholera virus, was accomplished using the same two types of conjugates. Specificity was confirmed by appropriate controls. The authors found immunofluorescence to be a convenient and sensitive method for determining an antigenic relationship between hog cholera and bovine viral diarrhea viruses.     

Hog Cholera Antibodies in Pigs Vaccinated with an Aujeszky-Vaccine Based on Antigen Produced in IB-RS-2 Cells

The cell line IB-RS-2 is confirmed to be persistently infected with hog cholera virus. Vaccination of pigs against Aujeszky’s Disease with an inactivated vaccine based on antigen produced on this cell line induced neutralizing antibodies against hog cholera virus.     

Glutaraldehyde Inactivation of Exotic Animal Viruses in Swine Heart Tissue

Glutaraldehyde, 0.2%, in a 1:100 (wt/vol) ratio, inactivated four animal viruses (foot-and-mouth disease, swine vesicular disease, African swine fever, hog cholera) in swine heart tissues during 11-day exposures at 22 to 26°C.     

Precipitation Reactions in Agar Between Swine Serum and Homologous Pancreas Extracts

Hyperimmune anti-hog cholera and nonimmune swine sera yielded approximately 50% more precipitation reactions in agar-gel diffusion tests with pancreas extracts from SPF noninfected swine than with extracts obtained from swine experimentally infected with virulent hog cholera virus. The pancreas-reacting property of swine serum was determined to be relatively heat stable, withstanding 68 C for 30 minutes. Of various swine serum fractions tested, the only one that reacted with pancreas extracts contained gamma, beta and alpha-globulins. In the absence of alpha-globulin, precipitation reactions were not observed. Sera of newborn SPF piglets, containing 50% alpha-2 globulin, formed more intense precipitation lines with swine pancreas extracts than were formed by the sera of their dams with the same extracts. The pancreas-reacting activity of swine sera was completely removed by absorption with pancreatic tissue. This property was not removed by absorption with guinea pig kidney, or beef, swine or human erythrocytes. Maceration of pancreatic tissue released reactive substances in a polydispersed form. This was demonstrated by the ability of almost all supernates and sediments from differential centrifugation of such preparations to form precipitation lines with swine sera. Reactive substances from swine pancreas were found to be relatively heat labile, being inactivated in one hour at 56C. No evidence was obtained in this study to indicate that the observed precipitation reactions were related to hog cholera virus and its corresponding antibody. The reactions are believed to have resulted from the interaction of protein-related substances present in normal swine pancreas with a relatively heat stable component, possibly alpha-globulin, in swine serum.     

Porcine leukocyte cellular subsets sensitive to African swine fever virus in vitro.

African swine fever virus infected most, if not all, of the macrophages (monocytes) and ca. 4% of the polymorphonuclear leukocytes from porcine peripheral blood. B and T lymphocytes, either resting or stimulated with phytohemagglutinin, lipopolysaccharide, or pokeweed mitogen, were not susceptible to the virus. All of the mitogens used inhibited African swine fever multiplication in susceptible cells. The number of virus passages in vitro and the virulence degree of the virus did not affect the susceptibility of porcine B or T lymphocytes to African swine fever virus.     

Efficacy of vaporized hydrogen peroxide against exotic animal viruses.

The efficacy of vapor-phase hydrogen peroxide in a pass-through box for the decontamination of equipment and inanimate materials potentially contaminated with exotic animal viruses was evaluated. Tests were conducted with a variety of viral agents, which included representatives of several virus families (Orthomyxoviridae, Reoviridae, Flaviviridae, Paramyxoviridae, Herpesviridae, Picornaviridae, Caliciviridae, and Rhabdoviridae) from both avian and mammalian species, with particular emphasis on animal viruses exotic to Canada. The effects of the gas on a variety of laboratory equipment were also studied. Virus suspensions in cell culture media, egg fluid, or blood were dried onto glass and stainless steel. Virus viability was assessed after exposure to vaporphase hydrogen peroxide for 30 min. For all viruses tested and under all conditions (except one), the decontamination process reduced the virus titer to 0 embryo-lethal doses for the avian viruses (avian influenza and Newcastle disease viruses) or less than 10 tissue culture infective doses for the mammalian viruses (African swine fever, bluetongue, hog cholera, pseudorabies, swine vesicular disease, vesicular exanthema, and vesicular stomatitis viruses). The laboratory equipment exposed to the gas appeared to suffer no adverse effects. Vaporphase hydrogen peroxide decontamination can be recommended as a safe and efficacious way of removing potentially virus-contaminated objects from biocontainment level III laboratories in which exotic animal disease virus agents are handled.     

Establishment of a Vero cell line persistently infected with African swine fever virus.

A Vero cell line persistently infected with African swine fever virus was established by infecting the cells in the presence of 10 mM NH4Cl (Vero-P cell line). The virus derived from the Vero-P cultures infected Vero cells, and virus titers were comparable to those obtained in Vero cells acutely infected with African swine fever virus. The structural proteins of the virus from Vero-P cells were similar to those of the virus produced in lytic infections. Virus production was low when the Vero-P cells were growing logarithmically and increased considerably in confluent cultures when lysis appeared in a fraction of the cell population.     

Effects of Oxyamylose and Polyacrylic Acid on Foot-and-Mouth Disease and Hog Cholera Virus Infections

Two interferon-inducing polycarboxylates were tested for antiviral activity on foot-and-mouth disease (FMD) virus infections in mice, guinea pigs, and swine. Polyacrylic acid, given intraperitoneally, had a protective effect on infection by FMD virus administered in the peritoneal cavity of mice and in the foot pad of guinea pigs. Chlorite-oxidized oxyamylose (COAM) was effective in mice at a dosage of 2 mg/kg. Swine were not protected against naturally transmitted FMD by 120 mg/kg of COAM nor by polyacrylic acid. Swine were not totally unresponsive to COAM since it delayed symptoms of hog cholera. Interferon was not detected in the serum of COAM-treated swine. With FMD virus, an example was found of activity of interferon inducers in experimental hosts and lack of activity in a natural host.     

Long-term persistent infection of swine monocytes/macrophages with African swine fever virus.

Long-term persistent infection was established in 100% of pigs (n = 19) experimentally infected with African swine fever virus (ASFV). Viral DNA was detected in peripheral blood mononuclear leukocytes (PBML) at greater than 500 days postinfection by a PCR assay. Infectious virus was not, however, isolated from the same PBML samples. In cell fractionation studies of PBML, monocytes/macrophages were found to harbor viral DNA during the persistent phase of infection. This result indicates that monocytes/macrophages are persistently infected with ASFV and that ASFV-swine monocyte/macrophage interactions can result in either lytic or persistent infection.     

The persistence of African swine fever virus in field-infected Ornithodoros erraticus during the ASF endemic period in Portugal

African swine fever (ASF) is an important disease of pigs and outbreaks of ASF have occurred in Europe on multiple occasions. To explore the period for which the European soft tick species Ornithodoros erraticus (Acari: Argasidae) is able to act as a reservoir of African swine fever virus (ASFV) after infected hosts are removed, we collected specimens from farms in the provinces of Alentejo and Algarve in Portugal during the endemic period and tested them subsequently using cell culture and experimental infection. We show that ticks from previously infected farms may contain infectious virus for at least five years and three months after the removal of infectious hosts. Furthermore, in two cases infectious virus was successfully isolated from ticks on restocked farms that had not yet suffered a re-emergence of disease. Experimental transmission to pigs was demonstrated in batches tested up to 380 days after an outbreak. These results clarify the epidemiological role of O. erraticus ticks in the persistence of ASFV in the field, provide additional evidence to support its role in the re-emergence of a sporadic outbreak of ASF in Portugal in 1999 and suggest that the current quarantine legislation and restocking advice when these ticks are present on the pig farm premises is appropriate.     

Use of the agar diffusion precipitation test in the diagnosis of hog cholera

The application of the agar diffusion precipitation (ADP) test for diagnosing hog cholera was investigated. The test used as antigen, pancreatic tissue from 272 pigs that had been inoculated with hog cholera virus. The test was positive for 13.5% of the animals that were sick for 4 days or less, 40% of those sick for 5 days, and 77% of those sick for 6 days or more. The test was positive for 13.5% of all animals that had been vaccinated with crystal violet-glycerol hog cholera vaccine and had been sick for at least 6 days after challenge inoculation. Titration of the virus in ADP-test-negative and ADP-test-positive pancreatic suspensions did not show a direct correlation between the infective virus particle and the precipitating antigen.     

Pathogenesis of classical swine fever: B-lymphocyte deficiency caused by hog cholera virus.

Hog cholera, also known as classical or European swine fever, is caused by hog cholera virus, a member of the genus Pestivirus. It is shown here that the end stage of lethal infection in the natural host is associated with a dramatic depletion preferentially of B lymphocytes in the circulatory system as well as in lymphoid tissues. Already at the onset of disease, viral replication in lymphoid tissues demarcates the germinal centers, and the viral genome remains localized to that site as the disease progresses even after morphologic disintegration of the follicular structure. A block in B-lymphocyte maturation by infection and destruction of germinal centers is discussed as a key event in the pathogenesis of acute, lethal hog cholera.