Naturally acquired antibodies to Bacillus anthracis protective antigen in vultures of southern Africa.

Sera from 19 wild caught vultures in northern Namibia and 15 (12 wild caught and three captive bred but with minimal histories) in North West Province, South Africa, were examined by an enzyme-linked immunosorbent assay (ELISA) for antibodies to the Bacillus anthracis toxin protective antigen (PA). As assessed from the baseline established with a control group of ten captive reared vultures with well-documented histories, elevated titres were found in 12 of the 19 (63%) wild caught Namibian birds as compared with none of the 15 South African ones. There was a highly significant difference between the Namibian group as a whole and the other groups (P < 0.001) and no significant difference between the South African and control groups (P > 0.05). Numbers in the Namibian group were too small to determine any significances in species-, sex- or age-related differences within the raw data showing elevated titres in four out of six Cape Vultures, Gyps coprotheres, six out of ten White-backed Vultures, Gyps africanus, and one out of three Lappet-faced Vultures, Aegypius tracheliotus, or in five of six males versus three of seven females, and ten of 15 adults versus one of four juveniles. The results are in line with the available data on the incidence of anthrax in northern Namibia and South Africa and the likely contact of the vultures tested with anthrax carcasses. It is not known whether elevated titre indicates infection per se in vultures or absorption of incompletely digested epitopes of the toxin or both. The results are discussed in relation to distances travelled by vultures as determined by new tracking techniques, how serology can reveal anthrax activity in an area and the issue of the role of vultures in transmission of anthrax.

Vibrio cholerae O1 outbreak isolates in Mozambique and South Africa in 1998 are multiple-drug resistant, contain the SXT element and the aadA2 gene located on class 1 integrons.

The characteristics of Vibrio cholerae O1 biotype El Tor, serotype Ogawa isolates from outbreaks of cholera in 1998 amongst migrant workers in the South African provinces of Gauteng and Mpumalanga, on the border of Mozambique, are reported. The isolates seem to have originated from the same clone since they are of two closely related BglI ribotypes. These ribotypes had a high similarity to ribotypes of V. cholerae O1 recently found in three South-east Asian countries. Isolates were resistant to furazolidone, streptomycin, sulfamethoxazole, trimethoprim and tetracycline. Only two isolates contained plasmids of 54 and 63 kb in size. PCR and DNA sequencing revealed that the chromosomally located resistance determinants present included an aadA2 gene cassette contained in a class 1 integron; the SXT element, which is a transposon-like element containing resistance genes; and the tetA gene. A co-transfer of chromosomal closely located genes encoding the SXT element and tetA was shown by mating experiments, PCR and pulsed-field gel electrophoresis analyses. Our study shows for the first time that multiple-resistant V. cholerae O1 isolates containing class 1 integrons and the SXT element were responsible for cholera outbreaks in Southern Africa. Studies are needed to determine the spread of this multiple-resistant O1 strain and further genetic details of the association of the SXT element, tetA and class 1 integrons, including their means of transfer.

Factors contributing to the emergence of Escherichia coli O157 in Africa.

In 1992, a large outbreak of bloody diarrhea caused by Escherichia coli O157 infections occurred in southern Africa. In Swaziland, 40,912 physician visits for diarrhea in persons ages >5 years were reported during October through November 1992. This was a sevenfold increase over the same period during 1990-91. The attack rate was 42% among 778 residents we surveyed. Female gender and consuming beef and untreated water were significant risks for illness. E. coli O157:NM was recovered from seven affected foci in Swaziland and South Africa; 27 of 31 patient and environmental isolates had indistinguishable pulsed-field gel electrophoresis patterns. Compared with previous years, a fivefold increase in cattle deaths occurred in October 1992. The first heavy rains fell that same month (36 mm), following 3 months of drought. Drought, carriage of E. coli O157 by cattle, and heavy rains with contamination of surface water appear to be important factors contributing to this outbreak.

In vitro activities of 14 antibiotics against 100 human isolates of Yersinia pestis from a southern African plague focus.

A limited repertoire of antimicrobial agents is currently in use for the treatment of plague. We investigated the in vitro activities of some newer antimicrobial agents against Yersinia pestis. Among the injectable agents tested, cefotaxime was the most active, and among the oral agents, both levofloxacin and ofloxacin were highly active, with MICs at which 90% of isolates are inhibited of < 0.03 microgram/ml. the susceptibilities to the ketolide RU004 and the penem faropenem warrant attention. The enhanced activities of quinolones against Y. pestis suggest that these agents should be further investigated for the treatment of human plague in the future.

Investigation of contaminated parenteral nutrition fluids associated with an outbreak of Serratia odorifera septicaemia.

An outbreak of fatal septicaemia caused by Serratia odorifera biotype 1 involved infants at several hospitals; the common vehicle of infection was contaminated parenteral nutrition fluid. The transfusate had been made up in a flexible film isolator system. The implicated organism was recovered from surfaces inside the isolator, despite routine decontamination procedures having been carried out shortly before. Our investigation into the origin of contamination revealed several shortcomings in the infusate compounding process. We noted deficiencies in cleaning and decontamination procedures, and in storage and sterility testing policies, but the origin and mechanism of the contamination were unclear. Withdrawal of parenteral nutrition products and revision of decontamination procedures terminated the outbreak. The efficacy of peracetic acid treatment of flexible film isolators, given the circumstances of this outbreak, may need further investigation. Regular training and assessment of admixture technicians is important.

Immune responses of two Mastomys sibling species to Yersinia pestis.

This study assessed the in vitro cell-mediated immune responses of Mastomys natalensis, with a diploid chromosome number of 2n = 32, and Mastomys coucha, with a diploid chromosome number of 2n = 36, to Yersinia pestis. Splenic mononuclear (MN) cells of uninfected M. natalensis proliferated in response to crude fraction 1 of Y. pestis and two subfractions derived from fraction 1 in vitro. Proliferation was dose dependent and followed the time kinetics of other well-known mitogens. Further characterization of the two fractions revealed similar protein profiles in sodium dodecyl sulfide-polyacrylamide gel electrophoresis and indicated a heat-stable protein of 25 kDa responsible for the mitogenic activity. No such response was observed with MN cells from M. coucha. The unresponsiveness of M. coucha-derived MN cells appears to be related to an inability to respond to Y. pestis organisms. The results may help explain the relative resistance and susceptibility of M. natalensis and M. coucha to Y. pestis infection.

Application of enzyme immunoassays for the confirmation of clinically suspect plague in Namibia, 1982.

An outbreak of plague occurred in Ovamboland, northern Namibia, late in 1982. Blood cultures, sera and blood clots were tested to obtain laboratory confirmations for clinically suspect cases of the disease. Isolation of the bacillus (Yersinia pestis) was attempted from blood cultures; sera were tested for antibody by passive haemagglutination (PHA) and enzyme-linked immunosorbent assay (ELISA). Sera and clots also were tested by ELISA for the specific F1 plague antigen. All the ELISA procedures were based on a monoclonal antibody to F1 antigen to ensure specificity.Thirty-eight cases were confirmed as plague: 50% by isolation, 34% by antibody responses, and 16% by the detection of antigenaemia. All isolates of Y. pestis were capable of producing F1 antigen, and significant antibody responses were observed in bacteriologically confirmed cases with paired sera. Patients who experienced sero-conversion had a higher IgM titre than IgG titre during the first nine days of hospitalization, while patients hospitalized for 17 or more days had IgG titres that were higher than the IgM titres. The relationship between IgM and IgG antibody titres is discussed with reference to identifying very recent infections. PHA titres increased and declined with IgM titres but were lower and more transient.ELISA procedures increased laboratory confirmations of plague by 23% above the numbers achieved using blood cultures and PHA tests alone. The ELISA to detect F1 antigen accounted for 86% of this increase by confirming cases where bacteriological isolation was not done. This ELISA did not replace the requirement for bacteriological isolation, since seven bacteraemic patients did not demonstrate antigenaemia.

Ecology of plague in Africa: response of indigenous wild rodents to experimental plague infection.

The Mastomys natalensis species complex, subdivided into genetically distinct species having diploid chromosome numbers 2n = 32 and 2n = 36, is a reservoir for several zoonoses including Lassa fever and plague. This report describes a study to determine whether these sibling species and three other rodent species have different potential as reservoirs for plague. It was found that M. natalensis (2n = 32) was significantly more resistant to experimental plague infection (50% survived inoculation with 120 000 Yersinia pseudotuberculosis subsp. pestis) than was M. coucha (2n = 36) (none of which survived doses of 190 Y. pseudotuberculosis subsp.pestis). In descending order of resistance were M. natalensis, Aethomys chrysophilus, M. coucha, Tatera leucogaster and A. namaquensis. No A. namaquensis survived inoculation of 10 or more plague bacilli.Previous reports on susceptibility to plague or other infections, which were based exclusively on findings in the universally distributed laboratory-bred Mastomys, are thus not necessarily applicable to the M. natalensis species as a whole but probably only to M. coucha. The Y. pseudotuberculosis subsp. pestis fraction-1 passive haemagglutination test appeared to be relatively insensitive in that only 5 out of 47 animals surviving experimental plague infection showed specific antibodies 6 weeks after challenge.The geographic distribution of human plague in southern Africa corresponds closely with that of the plague-susceptible species, M. coucha, while the resistant species, M. natalensis, predominates in areas where human plague has not been recorded. The role of A. namaquensis in the ecology of plague needs to be carefully studied and its possible importance in plague research should be investigated further.

Comparison of passive haemagglutination and enzyme-linked immunosorbent assay for serodiagnosis of plague.

Sera of 42 suspect plague cases from Ovamboland, Namibia, were examined. ELISA proved more sensitive than passive haemagglutination for the detection of F1 antibody and increased positive serodiagnoses of plague by 7-42%.

Ampicillin-resistant Haemophilus influenzae in Johannesburg.

Surveys carried out using a chromogenic cephalosporin test for beta-lactamase production (ampicillin resistance) among isolates of Haemophilus influenzae in Johannesburg have indicated an appreciable prevalence, especially among children seen at the new Johannesburg Hospital. Of type b strains recovered from these children, 10,9% were ampicillin-resistant. Three of the last 10 cases of serious systemic H. influenzae infections encountered at the Johannesburg Hospital were caused by beta-lactamase-producing strains, all of these having been acquired in the community rather than in hospital. These findings suggest that the optimal initial starting treatment for serious systemic or life-threatening H. influenzae infections should include chloramphenicol, either alone or at least in conbination with ampicillin or a similar compound.