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Electron microscopy study of the mode of growth of Pseudomonas pseudomallei in vitro and in vivo.

The mode of growth of Pseudomonas pseudomallei in culture media and in the lung tissue of infected humans and animals was studied using transmission electron microscopy (TEM) and scanning electron microscopy (SEM). In culture media, P. pseudomallei cells were seen to be entrapped in microcolonies within large amounts of intercellular fibrous material. The lung tissue of infected humans and animals showed that bacterial cells growing in lung tissue were surrounded by radially arranged fibres that constitute a very well defined glycocalyx structure. In the infected areas of the animal lung tissue, bacterial cell could be seen to have formed glycocalyx enclosed microcolonies that displaced host cell components, e.g. the nucleus of a phagocyte. The presence of bacteria in unusual locations indicated that effective phagocytosis was not occurring. The demonstration that cells of P. pseudomallei produce exopolysaccharide glycocalyces and form glycocalyx enclosed microcolonies in laboratory media and in lung tissue of infected humans and animals and the presence of bacteria in unusual locations contribute to a new understanding of the mechanism whereby this organism can cause persistent chronic infections.

Animals↗

Burkholder meningitis.

We report two cases of Burkholder meningitis. Case 1 presented with subacute lymphocytic meningitis, low sugar profile in CSF and positive culture for Burkholder. Case 2 presented with purulent meningitis that emerged during treatment of cryptococcal meningitis which, to our knowledge, has not been reported. Thus, Burkholder should be considered as one of the etiologic agents in this clinical setting.

Adult↗

[The immunogenicity and heterogeneity of Pseudomonas pseudomallei surface antigen 8].

Surface antigen 8, one of pathogenicity factors of P.pseudomallei and having an antiphagocytic action, contains glycoprotein with a mol. wt. of 200 kD and proteins with mol wt. of 25, 30 and 34 kD. According to its chemical structure, the carbohydrate part of antigen 8 is the homogeneous polymer of 6-d-D-mannoheptose, and its protein component is a collection of monomer proteins with mol. wt. of 12-120 kD. The consecutive fractionation of antigen 8 by gel and ion exchange chromatography has made it possible to isolate its individual fragments, differing by the collection of antigenic components, as well as by their antiphagocytic and protective activity. Experiments have shown that glycoprotein with a mol. wt. of 200 kD is an active immunosuppressive agent, while protein with a mol. wt. of 34 kD produces a pronounced immunogenic effect.

Animals↗

Evolution of cell-surface acid phosphatase of Burkholderia pseudomallei.

Acid phosphatase active fractions were obtained from cell-free extract, outermembrane fraction and culture filtrate of Burkholderia pseudomallei by column chromatography with sepharose 6B and DEAE cellulose. The comparison of the elution patterns of protein, sugar and enzymatic activity among these three components suggested that the enzyme is a glycoprotein evolving from premature proteins through glycosylation and that the enzyme is translocated during glycosylation from the cytoplasm to the outer membrane and finally excreted into the environment. When tunicamycin, a glycosylation inhibitor, was added to the culture, the peaks of sugar and enzymatic activity were lowered concomitantly leaving the protein peak unchanged in the elution pattern of the culture filtrate. The affinity of the bacterial surface to antienzyme sera was demonstrated by immuno-fluorescence microscopy.

Acid Phosphatase↗

Meliodosis in India.

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Antibodies, Bacterial↗

[Effect of TRA-system of plasmids RP4 and R68.45 on pseudomonas mallei virulence].

In Pseudomonas mallei, spontaneous mutants (Tra- mutants) of the plasmids RP4 and R68.45, losing the ability to transfer at conjugation are formed. The plasmids RP4 and R68.45 with Tra(+)-phenotype caused a decrease in P. mallei virulence for laboratory animals. At the same time, Tra- mutants of these plasmids do not affect P. mallei virulence. The insertion of DNA fragment of about 1900 bp into the plasmid transfer gene regions (tra-2-tra-3) gave rise to RP4 and R68.45 tra mutations detectable on examination.

Animals↗

Isolation rates of Burkholderia pseudomallei among the four regions in Thailand.

This study aimed to compare the isolation rates of Burkholderia pseudomallei among community-based hospitals located in the central, north, northeast, and south of Thailand. A questionnaire inquiring about the number of isolation of B. pseudomallei from various clinical specimens during 1994-95 were mailed to 141 community-based hospitals. Of these, 125 hospitals (88.6%) responded to the questionnaire. Microbiological laboratory was not available in thirty hospitals. Data from 95 remaining hospitals with capability to do bacterial culture showed that B. pseudomallei was never isolated in 49 hospitals. Eleven, 9, 19 and 7 hospitals where B. pseudomallei has been isolated, are located in the central, north, northeast and south of Thailand respectively. From these 46 hospitals, a total of 1,131 strains of B. pseudomallei were isolated from 407,263 specimens in 1994 and 1,165 strains from 440,541 specimens in 1995. However, the isolation was most frequent in northeastern hospitals, which accounted for 890 and 964 strains in 1994 and 1995 respectively while only 94, 76, 71 and 83, 75, 43 strains were simultaneously isolated during the 2-year period in those located in central, north and south respectively. The isolation rates of B. speudomallei in 1994 and 1995 were 4.2 and 4.1 per 1,000 clinical specimens in northeastern hospitals as compared to 1.1, 1.8, 1.1 and 1.1, 1.2, 0.7 in those located in central, north and south respectively. Ubon Ratchathani, Nakhon Ratchasima, Buri Ram, Khon Kaen and Udon Thani were the five provinces which exhibited the highest isolation rates as follows; 244, 150, 147, 127, 100 and 218, 128, 114, 119, 58, in 1994 and 1995, respectively. It was concluded that B.pseudomallei was most commonly isolated in the northeast of Thailand. Under-recognition of B. pseudomallei may prevail not only in other parts of Thailand but in some areas of the northeast as well.

Bacteremia↗