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W R Hess

Publications and source records attributed to W R Hess.

At least 55 records · Page 3Linked to original sources

Epitopic diversity of African swine fever virus.

African swine fever (ASF) is caused by an icosahedral cytoplasmic, double stranded DNA virus. In the acute form of the disease, pigs die from disseminated intravascular coagulation (DIC) with extensive damage of the free and fixed macrophage systems and the reticular epithelial cells of the thymus; mortality is virtually 100%. In recent years, subacute and chronic forms of ASF have become more prevalent in the field, especially in outbreaks occurring outside the continent of Africa, and virus isolated from these outbreaks have often been of lesser virulence. In pigs experimentally infected with such isolates, a number of immunopathological manifestations have been encountered, e.g. hypergammaglobulinemia associated with necrotizing pneumonia, persistent infection in the presence of ASF-specific antibodies, and lack of demonstrable virus neutralizing antibodies. Nevertheless, the immune systems of pigs that have clinically recovered have not been impaired by the infection. We suggest that the heterogeneous composition of the virus population in a given isolate may be one of the causes of the anomalous immune responses. When a number of biological markers, i.e., hemadsorption characteristics, plaque size, infectivity, virulence, antigenic determinants, and genomic structure, were used to characterize the virus clones derived from various ASF virus (ASFV) isolates, considerable heterogeneity was apparent. In the present investigation, 20 monoclonal antibodies (MAb), which specifically identified the 14 kDa viral protein within the cytoplasmic membrane of the infected cells, were used to determine epitopic differences among a number of virus clones derived from various isolates. All of the non-African isolates examined contained two epitopically different groups of virus clones, and the reaction profiles obtained were distinctly different from those obtained with the clones of an African isolate (Tengani). It was concluded that an ASFV isolate is composed of a biologically diverse virus population with distinctly different members which are only identified after cloning.

African Swine Fever↗

Potential arthropod vectors of African swine fever virus in North America and the Caribbean basin.

In an effort to identify arthropods that might serve as vectors and perhaps reservoirs of African swine fever virus (ASFV) if it were to enter the U.S.A., the blood-sucking insect Triatoma gerstaeckeri and four species of ticks of the genus Ornithodoros were established in colonies capable of reproducing in numbers sufficient to enable thorough studies to be made of their ASFV vector potentials. A nymphal stage of T. gerstaeckeri carried the virus for 41 days and retained it through one molt, but was unable to transmit it to susceptible pigs. Studies on O. coriaceus revealed that the species is able to harbor and transmit the virus for greater than 440 days, passing it trans-stadially from the first nymphal stage to the adult, sustaining it through at least four molts. Trans-ovarial passage was not demonstrated and nearly 40% of the ticks died, apparently, of the ASFV infection. O. turicata collected in Florida was also found to be capable of becoming infected with ASFV and transmitting it by bite to susceptible pigs. O. puertoricensis collected during the ASF eradication programs in the Dominican Republic and Haiti was not only readily infected experimentally, but it was also able to transmit the virus trans-stadially and trans-ovarially. However, ASFV was not isolated from any of the 350 O. puertoricensis collected in the Dominican Republic and Haiti. O. parkeri from a long-established laboratory colony were able to carry the virus through at least one molt, but they were unable to transmit it to susceptible pigs.

African Swine Fever↗

Identification of African swine fever viral antigens in the hemolymph of soft ticks (Argasidae: Ornithodoros) by the immunodot blot test.

The immunodot blot test was used to identify African swine fever virus (ASFV) antigens in the hemolymph from soft ticks (Ornithodoros coriaceus) fed on ASFV-infected pigs. The immunodot blot test was sensitive and specific for ASFV antigens and has potential field application. Hemolymph from field-collected ticks can be screened for ASFV and a variety of other tick-borne pathogens using this test.

African Swine Fever Virus↗

Diversity of African swine fever virus.

An African swine fever virus is an heterogeneous population, consisting of clones having different biological characteristics in respect to hemadsorption, virulence, infectivity, plaque size, and antigenic determinants. The following observations were made: Nonhemadsorbing virus (NHV) have been segregated from field isolates from Haiti (HT-1) and a bone marrow- and buffy coat-passaged Portuguese isolate (L'60BM89BC1) and appear as a major, minor, or equal mixture with hemadsorbing viruses in the virus population. Biological characteristics of the virus inoculated into pigs often differed from viruses isolated later from the same pigs. Virulence and nonhemadsorbing characteristics of isolated clones were genetically stable. The lethal effect of 2 NHV clones of L'60BM89BC1 virus was dose-dependent; small doses of virus induced immunologic deaths or recoveries from the clinical disease in pigs, and large doses induced acute deaths. The NHV of Lisbon isolate of 1960 (L'60) and HT-1 isolate share the same antigenic determinants for inducing protection. Tengani isolate contained clones of distinctly different antigenic determinants, not shared by L'60 or HT-1 isolate that enabled it to overcome the protection induced by the other clones. Passaging of an African swine fever virus isolate in pigs or cell cultures may readily alter the proportions of the different clones in the population and thereby change its overall characteristics. A new virus population with atypical hemadsorption was found in HT-1 field isolate and L'60BM89BC1 virus.

African Swine Fever↗

Virulence in African swine fever: its measurement and implications.

A method of measuring and expressing the virulence of African swine fever virus in numerical terms was developed. Seventeen viruses (13 hemadsorbing and 4 nonhemadsorbing) were tested and classified into 3 groups: highly infectious and highly virulent, highly infectious and moderately virulent, and slightly infectious and slightly virulent. This classification was based on the number of 50% hemadsorption unit (HA50) or TCID50 required to produce 1 LD50 for swine, the number of HA50 or TCID50 required to produce one 50% pig infectious dose (PID50), and the number of PID50 required for each LD50. The virulent virus (group 1) required less than or equal to 10 virus units (HA50 or TCID50) for 1 PID50 and LD50 (highly infectious and highly lethal), respectively, and had a ratio of 1.0 for PID50/LD50, ie, all infected pigs died from acute African swine fever. Tengani, L'60, and DR-I isolants and nonhemadsorbing viruses of Haiti-1 isolant belong to this group. The moderately virulent virus (group 2) required less than 10 virus units for 1 PID50 and 20 to 562 virus units for 1 LD50 (highly infectious and moderately lethal), respectively, and recoveries from the clinical disease and immunologic deaths were frequent. Madrid '75, Haiti-1, DR-II, and BR-I isolants belong to this group. The slightly virulent virus (group 3) required 56 to 10,000 virus units for 1 PID50 and 56,200 to 3,120,000 virus units for 1 LD50 (slightly infectious and less lethal).(ABSTRACT TRUNCATED AT 250 WORDS)

African Swine Fever↗

New method of antibody detection by indirect immunoperoxidase plaque staining for serodiagnosis of African swine fever.

An indirect immunoperoxidase plaque-staining method was developed for detecting antibody to African swine fever virus infection. In both sensitivity and specificity, the test was comparable to indirect immunofluorescence. Because it has all of the desirable features of the indirect immunofluorescence test and may also be readily used for testing large numbers of sera, the indirect immunoperoxidase plaque-staining method can be used as a single and final serodiagnostic test in a large-scale survey of African swine fever. The indirect immunoperoxidase plaque-staining method should be applicable to other viruses that can be adapted to and grown in cell cultures.

African Swine Fever↗

Replication of African swine fever virus in cell cultures.

Infection-specific, nonstructural, African swine fever virus antigens, as visualized by the immunofluorescence technique, appeared as fine stipplings, evenly distributed in the cytoplasm and nucleus of Vero cells and in monocytes by postinoculation hour (PIH) 3. Cytoplasmic inclusion bodies (IB) appeared at PIH 4 and continued to increase in size up to PIH 8. The viral DNA was solely synthesized within the IB of infected monocytes, as evidenced by an autoradiographic chase experiment; maximum synthesis occurred at PIH 5. The 2,300-S and 2,500-S (sedimentation coefficient) structural viral antigens were visualized in the IB at PIH 5 and 6, respectively. The infective new progeny of African swine fever virus were formed between PIH 7 and 8, and the cell surface viral antigens were produced between PIH 8 and 9. Free virus was released between PIH 10 and 11. Infected cells detached from the glass surface from PIH 13. The 5-iodo-2'-deoxyuridine interfered with the formation of infective virus, but not interfere with the production of viral antigens.

African Swine Fever Virus↗

Sedimentation coefficient of African swine fever virus.

The sedimentation coefficient of the infective unit of African swine fever in tissue culture harvest fluids was measured in a preparative ultracentrifuge. The boundary locator method used also permitted making an estimate of heterogeneity. The sedimentation coefficient ranged from 3,000 to 8,000 Svedberg units, representing many classes of infective particles. Electron microscopy on culture fluids from infected cells showed many kinds of virus-containing units. Sucrose-CsCl gradient centrifugation was used to concentrate and to purify (partly) African swine fever virus for analytical ultracentrifugation. The optical patterns of the physical particles revealed a range of coefficients from 1,800 to 3,200 Svedberg units in tris-buffered saline solution at 20 C and buoyant densities from 1.19 to 1.24 g/ml in CsCl. The disparity of these values from those obtained by preparative ultracentrifugation indicates a change in the virus structure or a selection of viral populations on purification (or both).

African Swine Fever Virus↗

Experimental transmission of African swine fever virus by Ornithodoros coriaceus, an argasid tick indigenous to the United States.

Three ticks, indigenous to the United States, were assessed for their ability to maintain and transmit African swine fever virus (ASFV). Amblyomma americanum and A cajennense adults and nymphs maintained virus for 4 to 7 days after engorging on infected swine blood; however, virus was not carried through the subsequent molt of nymphs and it was not transmitted vertically to the eggs and larvae of the infected females. On refeeding of the ticks, infection was not transmitted to healthy swine. Ornithodoros coriaceus adult females also did not show transovarial transmission of two strains of ASFV. Nymphs, however, maintained virus for 77 to 118 days through a molting stage and transmitted one strain of the virus (Lisbon 60) to healthy swine.

African Swine Fever↗

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.

African Swine Fever Virus↗

African swine fever: microplaque assay by an immunoperoxidase method.

A microplaque assay for Vero cell-adapted Lisborn '60 strain of African swine fever virus (L'60-uncloned) and a large plaque-forming strain cloned from the L'60-uncloned strain was developed by an immunoperoxidase method. The immunoperoxidase method can be used to stain microplaques of 3 days after inoculation, whereas the conventional plaque assay requires 5 to 7 days to develop visible plaques. A linear relationship between viral concentration in the inoculum and plaque numbers was observed. Viral titers obtained by both microplaque assay and conventional plaque assay were comparable, and both methods were reproducible and reliable. The viral titer obtained by either one of the plaque assay methods was approximately 0.9 log10 lower than that obtained by the hemadsorption test.

African Swine Fever Virus↗

Cellular immunity demonstrated in pigs infected with African swine fever virus.

Twenty-two pigs infected with African swine fever virus (ASFV) were used to demonstrate delayed hypersensitivity (DH) in vitro by the leukocyte migration-inhibition test. The results indicated that ASFV-infected pigs developed DH against ASFV antigen (ASF antigen) as early as 20 days after inoculation, and the presence of viremia did not interfere with the leukocyte migration-inhibition test. Three ASFV-infected pigs that were also sensitized to mycobacterium developed DH against both ASF antigen and mycobacterium. The conclusion was that the cellular immune system is not impaired by ASFV infection in pigs.

African Swine Fever↗

T and B lymphocytes in porcine blood.

Two subpopulations of lymphocytes in the peripheral blood of young, conventionally reared Tamworth pigs had the following cell-surface markers analogous to those of human lymphocytes; (1) T cells--receptor(s) for ovine and caprine erythrocytes, permitting spontaneous formation of rosettes and the presence of the thymus-derived antigen; and (2) B cells--membrane-associated immunoglobulin and receptor(s) for complement. The T cells could be identified by either the erythrocyte rosettes or the cytotoxicity test, and the B cells could be identified by either the erythrocyte-antibody-complement rosette test or the immunofluorescence test for surface immunoglobulin. Because of simplicity, the cytotoxicity and immunofluorescence tests are recommended. The cell losses in ficoll-diatrizoate gradient centrifugation were studied with 9 blood samples. Of the total T and B cells, 71.7% were in fraction 1, and 28.3% were in erythrocyte fraction 2. Recovery rates of total leukocytes varied widely in relation to leukocyte numbers present in the original blood; the range was between 52.6 and 93.8%. Moreover, evidence did not indicate that either T or B cells were selectively sedimented in fraction 2. Analysis of the lymphocyte-rich fractions (fraction 1) from 37 blood samples by ficoll-diatrizoate gradient centrifugation indicated the following: (1) Approximately 2.7 X 10(6) T cells and 0.9 X 10(6) B cells could be obtained in the fraction from 1 ml of peripheral blood; (2) T and B cell populations were independently distributed; hence, no definite ratio between T and B cells applied to an individual animal; (3) approximately 20% of the lymphocytes did not have markers as either T or B cells; and (4) an average of 26% cells were peroxidase-positive leukocytes, chiefly monocytes.

Animals↗