Comparative tests for detection of plague antigen and antibody in experimentally infected wild rodents.

The enzyme-linked immunosorbent assay (ELISA) was compared with other standard tests for detection of plague (Yersinia pestis) antibody and antigen in multimammate mice (Mastomys coucha and M. natalensis) which were experimentally infected and then killed at daily intervals postinoculation. For detection of antibody in sera from M. natalensis, the immunoglobulin G (IgG) ELISA was equivalent in sensitivity to passive hemagglutination and more sensitive than the IgM ELISA and complement fixation. Antibody was first detected on postinfection day 6 by all four tests, but IgM ELISA titers had declined to undetectable levels after 8 weeks. For detection of fraction 1 Y. pestis antigen in rodent organs, the ELISA was less sensitive than fluorescent antibody but more sensitive than complement fixation or immunodiffusion. Plague fraction 1 antigen was detected in 16 of 34 bacteremic sera from M. coucha and M. natalensis. The threshold sensitivity of the ELISA was approximately 10(5) Y. pestis per ml.

Comparative pathogenicity and antigenic cross-reactivity of Rift Valley fever and other African phleboviruses in sheep.

Homologous and heterologous haemagglutination-inhibition (HAI), complement-fixation (CF), immunodiffusion (ID) and mouse neutralization tests were performed with the Lunyo (LUN) and a Zimbabwean strain of Rift Valley fever (RVF) virus, the prototype and a South African strain of Arumowot (AMT) virus and prototype strains of Gordil (GOR), Saint-Floris (SAF) and Gabek Forest (GF) viruses, using immune mouse ascitic fluids prepared against these viruses. Reactions of identity occurred in all tests between LUN and the Zimbabwean strains of RVF and between the two strains of AMT virus. Otherwise, cross-reactions occurred between all the phleboviruses in HAI tests, while reactions in CF, ID and neutralization tests were monospecific for virus serotypes, except that weak cross-reaction occurred between GOR and SAF viruses in CF and ID tests. Four sheep infected subcutaneously with the Zimbabwean strain of RVF virus developed transient fever, viraemia, leucopaenia, relative thrombocytopaenia, haemoconcentration and raised serum enzyme levels, which indicated that the sheep had developed necrotic hepatitis. Disseminated focal necrotic hepatitis was confirmed in a sheep killed for examination on day 4 post-infection. The other three sheep recovered uneventfully after only mild depression and anorexia. Groups of three sheep infected with SAF, GOR, AMT and GF viruses had no demonstrable viraemia or other sign of infection or illness, except that the sheep infected with AMT developed mild fever lasting less than 24 h. Antibody responses were monitored at intervals over a period of 24 weeks in all sheep by homologous and heterologous HAI, CF and cell culture neutralization (CPENT) tests. Homologous antibody responses were marked in the RVF-infected sheep and their sera cross-reacted strongly in HAI tests with antigens of the other viruses. The sera of the RVF-infected sheep cross-reacted less markedly in CF and CPENT tests. Homologous antibody responses were poor in all the sheep infected with phleboviruses other than RVF, and the cross-reactivity of their sera for RVF antigen or virus was negligible. All sheep were challenged with RVF virus 48 weeks after their initial infection. The sheep which had originally been infected with RVF virus were immune and developed neither fever nor viraemia. All other sheep developed fever, viraemia and antibodies to RVF virus. It was concluded that the African phleboviruses, other than RVF, are unlikely to cause disease in livestock or to induce antibodies which could cause confusion in the diagnosis of RVF.

Experimental plague infection in South African wild rodents.

Susceptibility studies were undertaken to determine the response of some South African wild rodent species to experimental plague (Yersinia pestis) infection. A degree of plague resistance was found in three gerbil species captured in the plague enzootic region of the northern Cape Province, these being the Namaqua gerbil, Desmodillus auricularis, (LD50 1 X 10(6) organisms), the bushveld gerbil, Tatera leucogaster, (LD50 9.1 X 10(5)) and the highveld gerbil, T. brantsii (LD50 4 X 10(2)). Animals from a population of the four-striped mouse, Rhabdomys pumilio, captured in the plague area of Port Elizabeth, proved moderately resistant to experimental plague infection (LD50 1.3 X 10(4)) while those from another population of the same species captured in a plague-free area of the Orange Free State were extremely susceptible (LD50, 5 organisms). The response of both populations however was a heterogeneous one. Marked differences in susceptibility were also found between two populations of multimammate mice, Mastomys natalensis (2n = 32) although both originated from areas outwith the known distribution of plague in southern Africa. The 50% infectious dose was relatively high in T. leucogaster (3.2 X 10(2)) and D. auricularis (1.7 X 10(3)), but was low (2-16 organisms) in the other rodent species tested. The plague antibody response, determined by enzyme-linked immunosorbent assay (ELISA), was extremely short-lived in T. leucogaster, only 10% of inoculated animals remaining seropositive at low titres after 11 weeks. Antibodies persisted for only slightly longer in the sera of T. brantsii which were reinoculated with 2 X 10(3) plague organisms 6 weeks after initial challenge. The demonstration of the existence of both susceptible and resistant populations of R. pumilio and M. natalensis indicates that these species must be considered as potential plague reservoir hosts in parts of South Africa. The results suggest that resistance to plague infection in previously epizootic hosts in the northern Cape Province such as Tatera sp. and D. auricularis has arisen through continual selective pressure of the organism. If the findings are applicable to gerbil populations in other plague enzootic regions of South Africa it is probable that acquired plague resistance has been responsible for the absence of gerbil epizootics and consequently for the dramatic decline in human plague outbreaks in South Africa since 1950.

A comparison of serological techniques for plague surveillance.

Sera collected from 691 dogs and 231 rodents in South Africa were tested in parallel by the passive haemagglutination (PHA) and enzyme-linked immunosorbent assay (ELISA) for antibodies to the Fraction 1 antigen of the plague bacillus (Yersinia pseudotuberculosis subsp. pestis). There was a high degree of correlation between the tests, both in over-all numbers positive and in serum titres. 42 dog sera were positive by PHA and 41 by ELISA, 40 sera being positive by both tests. However, using ELISA the numbers of non-specific reactors in dog sera were cut by almost two thirds compared to PHA. The results of tests on rodent sera were identical: only one rodent was positive in both tests. The findings suggest that the ELISA test may have advantages over PHA for large scale serological surveillance.

Studies on plague in the eastern Cape Province of South Africa.

Investigations were carried out in the area of a human plague outbreak in March 1982 at Coega in the eastern Cape Province of South Africa. Trapping revealed that rodent populations were high owing primarily to a population increase of the four-striped mouse, Rhabdomys pumilio. Flea populations were low and had been declining in the eastern Cape Province since October 1981. The results of a serological survey from March to October 1982 showed haemagglutinating antibody to Yersinia pestis in 0.35% of 5938 dog sera and 0.53% of 1132 small mammal sera. Plague antibody was detected in two small mammal species, R. pumilio and the vlei rat, Otomys irroratus. There was thus evidence that a small, limited rodent epizootic preceded the human plague cases and it appears that at least one human case was contracted from an infected domestic cat. The extent of the enzootic area was confined to a coastal belt north of Port Elizabeth and showed a close correlation with river courses. It was concluded that the Port Elizabeth-Uitenhage focus was in an inter-epidemic phase and that the human plague outbreak was an isolated incident.