- Volume 48, Issue 6, 1999

Two types of fimbriae, designated aggregative adherence fimbria I and II (AAF/I and AAF/II) have been described in enteroaggregative Escherichia coli (EAEC) strains. These fimbriae mediate the aggregative pattern of adherence (AA) to epithelial cells. The genes encoding the structural subunit of each fimbria have been designated aggA and aafA, respectively. The prevalence of these genes was investigated in 155 faecal EAEC isolates that displayed AA to HeLa cells and were isolated from children in Sao Paulo, Brazil. Hybridisation assays with aggA and aafA sequences showed that 9.7% of these isolates carried aggA, 3.9% aafA, and none hybridised with both sequences. Of the 78 isolates displaying AA that reacted with the previously described EAEC probe (CVD432), 19.2% and 7.7% hybridised with the aggA and aafA probes, respectively. None of the 77 isolates displaying AA but lacking the EAEC probe sequence hybridised with either probe. These results clearly indicate that additional factors are involved in the AA phenotype in these EAEC strains.

BALB/c and severe combined immunodeficient (SCID) mice were inoculated intraperitoneally with Mycobacterium avium and the numbers of cfu were monitored for 70 days in spleen, liver, lung, kidney, brain and peritoneum. While BALB/c mice formed typical granulomas and controlled bacterial growth in organs, a delay in development of lesions and a modest containment of infection were observed in SCID mice. In the spleen of BALB/c mice, in which bacterial growth was contained, macrophages (M⊘) and natural killer (NK) cell numbers increased ≥4.2 times and T- and B-cell numbers increased ≥1.8 times after 42 days of infection; conversely, a low recruitment of mononuclear cells was observed in the spleen of SCID mice, where M. avium proliferated efficiently. Unlike visceral organs, a pronounced decrease in the number of cfu was observed in the peritoneum of BALB/c mice, concomitantly with a ≥31.7-fold increase in M⊘ and NK cells and a ≥9.1-fold increase in T and B cells. In the peritoneum of SCID mice only a bacteriostatic effect was observed despite a ≥56.7-fold increase in M⊘ and NK cells and a ≥22.3-fold increase in T and B cells. These results suggest that while an intact immune response can efficiently control M. avium infection in the spleen and peritoneum of BALB/c mice, cells of the innate immune system such as M⊘ and NK cells play a role in the containment of bacterial growth in the peritoneum, but not spleen, of SCID mice.

Many methods have been described for the detection of methicillin-resistant Staphylococcus aureus (MRSA), but the homogeneous or heterogeneous expression of methicillin resistance affects the reliability of those methods. This study demonstrates that close association between methicillin-susceptible S. aureus (MSSA) and MRSA strains in the host colonisation site can present additional problems for the detection of MRSA in clinical laboratories, which may contribute to failure in the control of MRSA infection in hospital. Worse, this association may also account for the emergence of MRSA during antibiotic therapy.

Enterobacter sakazakii, designated a unique species in 1980, has been implicated as the causative organism in a rare but severe form of neonatal meningitis. Dried infant formula milk has been identified as a potential source of the organism. E. sakazakii isolates from dried infant formula available in Canada and clinical isolates obtained from Canadian hospital culture collections were characterised by phenotypic (biotype and antibiograms) and genotypic (ribotyping, random amplification of polymorphic DNA and pulsed-field gel electrophoresis) methods. Three biotypes and four antibiogram patterns were observed in the 18 isolates examined. Ribotyping with the Dupont Ribopiinter™ microbial identification system divided the 18 isolates into 10 ribotypes. Three isolates from the same hospital had indistinguishable ribotyping patterns although each was isolated in a different year, as did three food isolates from one company. Pulsed-field gel electrophoresis (PFGE) and random amplification of polymorphic DNA (RAPD) profiles indicated minor differences between the isolates that were indistinguishable by ribotyping. PFGE (with the restriction endonucleases Xba1 and Spel) and RAPD gave discrete patterns that enabled easy comparison of E. sakazakii isolates, with a high degree of discrimination. The discriminatory index showed RAPD and PFGE were shown to be the most discriminatory typing schemes for E. sakazakii, followed by ribotyping, biotyping and antibiograms.

In a hospital-based study, stool samples from 2095 patients of all ages were examined for different fungal, protozoal and bacterial enteropathogens over a period of 2 years (July 1994-June 1996). Cryptosporidium was detected in 151 specimens (7.2%) and was the third commonest pathogen found. The highest prevalence of this organism was in the group aged 16-45 years and during the rainy months (July-Oct.). Diarrhoea caused by the protozoon was of mild to moderate severity and features of dysentery were absent. Amongst other enteropathogens, Candida albicans was the most frequently isolated, followed by enteropathogenic and enterotoxigenic Escherichia coli, Salmonella spp., Campylobacter jejuni, Entamoeba histolytica, Giardia duodenalis (lamblia), Shigella spp., Vibrio cholerae and Aeromonas spp.

This report describes extraction of a new native antigen fraction from Mycobacterium tuberculosis without massive degradation of proteins by Triton X-100. The Triton X-100 solubilised protein (TSP) antigen showed a characteristic antigen profile and reproducible extraction pattern. To characterise the nature of their composition, the TSP antigen was fractionated by Triton X-114 phase partitioning. The TSP antigen contained a variety of lipids and glycoconjugates as well as diverse proteins. Most proteins were partitioned into the aqueous phase during phase fractionation, whereas non-protein molecules and lipoproteins were recovered in the detergent phase. The lymphoproliferative responses to the TSP aqueous fraction in healthy tuberculin reactors were significantly higher than those to the purified protein derivative (PPD) and unfractionated TSP. In contrast, the antibody responses to TSP aqueous fraction in tuberculosis patients showed weak reactivity. This study suggests that the TSP aqueous fraction can be used as a T-cell antigen associated with protective immunity against tuberculosis.

Two fatal cases of Campylobacter jejuni septicaemia in patients with AIDS were characterised by severe HIV-related immunodeficiency, negative stool cultures and presentation during hospitalisation, developing a clinical picture of fulminant septic shock despite therapy with appropriate antibiotics. Campylobacter spp. are important opportunist pathogens in HIV disease and may cause a septicaemic illness in the absence of enteric disease.

A uropathogenic strain of Proteus mirabilis was grown in vitro in human and mouse urine and brain-heart infusion broth (BHIB) and in vivo in subcutaneous open chambers (SOC) in mice, intraperitoneal diffusion chambers (IPC) in rats and by ascending urinary tract infection in mice in order to compare growth pattern, cellular differentiation and expression of virulence factors. Although the growth rate was slower in vivo than in vitro, the extent of growth was similar after 24 h. P. mirabilis differentiated into filamentous swarmer cells in all in-vitro culture conditions, but no filamentous cells were observed in either of the in-vivo chamber models. Transurethrally infected mice showed a rapid release or loss of filamentous cells and these could not be seen in kidney or bladder homogenates 7 days after infection. Bacteria showed increasing haemagglutination titres for fresh and tanned red blood cells after subculturing in BHIB, but bacteria grown in vivo did not show haemagglutination. An increasing resistance to normal serum was found when bacteria were grown in vivo. Significant haemolytic activity was detected with bacteria grown in BHIB and IPC, but almost no activity was found when bacteria had grown in urine. These findings improve the understanding of the role of P. mirabilis uropathogenic virulence factors in vivo.

This report is an overview of poliomyelitis surveillance in Tunisia from 1991 to 1996. In all, 2088 stool specimens, collected from 152 acute flaccid paralysis (AFP) cases and from 1747 of their healthy contacts were investigated. Virus isolation was done systematically in RD and HEp-2C cell lines and isolated viruses were typed by seroneutralisation as polioviruses or non-polio enteroviruses. Poliovirus isolates were analysed systematically for their wild or vaccine-related origin by two methods - one based on antigenic differences and one on genetic differences between strains. All type 2 polioviruses were vaccine-related and most wild viruses belonged to polio serotype 3. Wild polio type 3 viruses were detected in 1991 and 1992 in six cases of paralytic polio. A silent circulation of wild polio 1 and wild polio 3 was detected in 1994. No wild virus was detected in Tunisia from 1995 onwards. Wild polioviruses were sequenced and compared with Tunisian wild strains isolated during the 1980s, as well as other genotypes from the international database. These investigations revealed a single Tunisian polio 3 genotype that has been circulating from 1985 to 1994 and two different polio 1 genotypes. These results reflect effective control strategies within the country and contribute to the improvement of the polio eradication programme effectiveness at national and global levels.