- Volume 47, Issue 6, 1998

The Ehrlichieae are gram-negative obligately intracellular bacterial pathogens. They can be divided into at least three genogroups on the basis of 16S rRNA gene sequences, but are also classified by target cell specificity. A group of granulocytic ehrlichiae primarily infect neutrophils and fall into genogroup II. The granulocytic ehrlichiae are subdivided by their target hosts, i.e., Ehrlichia phagocytophila in cattle and sheep, E. equi in horses, and the agents of human (HGE) and Ilama (LGE) granulocytic ehrlichioses. However, these subdivisions may give a false impression, as all these species are closely related both antigenically and on the basis of 16S rRNA operon sequence. In addition, cross-species transmission can occur naturally or by experimental infection. The vectors for these granulocytic ehrlichiae are hard-bodied ixodid ticks, and the reservoir hosts are probably wild rodents, deer and sheep. In each host, this illness presents as a febrile disease which can be followed by immunosuppression leading to secondary infections.

A total of 2800 sputum samples referred for mycobacterial investigation was examined by both continuous automated mycobacterial liquid culture (CAMLiC) and conventional Loewenstein-Jensen culture (LJC). The CAMLiC system was more sensitive than LJC, detecting 188 (98.4%) of 191 of all mycobacteria found by one or both methods compared to 150–162 (78.5–84.8%) found by LJC (the range for LJC takes into account all potential ‘missed positives’ due to contamination). Figures for Mycobacterium tuberculosis complex (MTBC) organisms specifically were 133 (98.4%) of 135 for CAMLiC and 115–122 (85.2–90.4%) for LJC. Detectable growth of MTBC organisms in CAMLiC occurred at a mean of 13.4 days after inoculation (range 3–32; SD 6.49; mode 8 days); 65.4% of such isolates were detected within 14 days and 87.2% within 21 days. In 73 instances the MTBC status of the isolate was defined by gene probe on the day of growth detection. Sufficient biomass for valid gene probe assay was always present. The speed, sensitivity and labour-sparing technology (no manual intervention is necessary before identification or discard) of CAMLiC make it possible for many laboratories to approach the Centers for Disease and Prevention (CDC) standard for culture and identification in mycobacteriology without resort to direct DNA detection techniques and at a much lower cost.

The development of Pneumocystis carinii infection in immunosuppressed rats is important not only in understanding the infection, but also as a source of P. carinii antigen for use in diagnostic serological tests. The aims of this study were to monitor the progression of P. carinii infection in the Sprague Dawley rat model and then determine parameters that indicate the maximum production of P. carinii antigen. Seventeen Sprague Dawley rats were killed at intervals up to 9 weeks after the start of immunosuppressive therapy. The progression of P. carinii lung infection was observed by Giemsa staining of lung imprints and by a hemi-nested polymerase chain reaction (PCR). Body weight, food and water intake and the appearance and activity of the rats were measured daily. Seven control rats were kept under the same conditions. P. carinii infection was detected in the lung 2 weeks after immunosuppression by hemi-nested PCR and after 3 weeks by Giemsa staining. No P. carinii DNA was detected in any of the blood samples. Rats with moderate or severe lung infection had been immunosuppressed for ≥ 6 weeks. Body weight was significantly greater in control rats than in the immunosuppressed rats. Six weeks of immunosuppression was used as a cut-off to determine measurements to identify those rats with moderate or severe infections in their lungs. A combination of >34% body weight loss at 6 weeks after immunosuppression and the condition of the animals with scores ≤ 9 used in conjunction with duration of immunosuppression may be useful to maximise the yield of P. carinii infection from individual rats.

Monoclonal antibodies (MAbs) that inhibit adhesion of Helicobacter pylori to human gastric cancer (MKN45) cells were established to clarify the mechanism of adhesion of H. pylori. Of 53 hybridoma clones screened by the primary inhibition assay for adhesion, MAb A20 of IgM class was selected on the basis of both its reactivity to whole cells of H. pylori by ELISA and its inhibitory effect on adhesion of H. pylori. The adhesion of H. pylori strain TK1029 to MKN45 cells was inhibited by MAb A20, depending on the concentration of the MAb. The MAb recognised the surface antigen, lipopolysaccharide (LPS) of H. pylori, suggesting that LPS is associated with adhesion of H. pylori to human gastric epithelial cells.

This study confirmed that Feinberg and Whittington medium was suitable for the cultivation and detailed study of the growth cycle of two clinical strains of Trichomonas vaginalis under anaerobic conditions. Both strains showed a similar growth pattern characterised by early but slow growth, extended duration of the logarithmic phase and limited survival never exceeding 144 h. Duration of survival and growth rate were inversely proportional to the inoculum density. Growth rate was pH dependent; pH values in the range 6.9–6.5 delayed the initiation of growth of T. vaginalis for at least 48 h. On the other hand, pH values of 6.4–4.5 were indifferent or slightly favourable for growth during the logarithmic and survival in the early decline phase. Normal saline and Ringer’s solution exerted an early and progressively lethal effect on trichomonads and led to the disappearance of protozoa suspended in them in 150 min. In general, these in-vitro results shed light on some aspects of the biology of T. vaginalis and contribute to a better understanding of the epidemiology and clinical manifestations of the infection.

During the period 1979–1991, Salmonella Typhimurium DT 204c was the cause of a major epidemic of salmonellosis in calves in the UK. Plasmid profile analysis of DT 204c isolates from England and Wales commenced in 1986 and isolates from all subsequent incidents were examined by this technique. Forty-three different plasmid profile types (PPTs) were detected, of which the commonest, designated type E, constituted 44.6–80.2% of the annual incidents during the study period. Some PPTs, e.g., F and P, were detected throughout most years of the study, whereas PPTs O and 6 persisted for short periods. Until 1984, most isolates were resistant to neomycin, but the subsequent predominant PP type E was sensitive to this antibacterial agent. It was concluded that during the epidemic there was an evolution of new genotypes, of which only some persisted; again, antibacterial resistance genes may be acquired or lost. The study demonstrated the value of PP typing for epidemiological studies.