Penelope R. Sisson

PHAGOCYTOSIS AND KILLING OF BACTERIA IN AEROBIC AND ANAEROBIC CONDITIONS

Phagocytosis and intracellular killing of several facultative anaerobes were observed in aerobic and anaerobic conditions. The presence of anaerobes affected these processes. Bacteroides asaccharolyticus and B. fragilis were killed by phagocytes only at bacterial concentrations less than 1 x 10(7) cfu/ml; at higher concentrations of the anaerobes, killing of B. fragilis and concomitant facultative anaerobes was inhibited. This effect appeared to be due to an interaction, in appropriate reducing conditions, between anaerobe and serum, which allowed engulfment of organisms by phagocytes but markedly impaired intracellular killing.

Direct milk excretion of Campylobacter jejuni in a dairy cow causing cases of human enteritis.

Consumption of milk contaminated with Campylobacter jejuni has been described as a cause of human enteritis. Although faecal contamination of milk with the organism has frequently been described, direct milk excretion of Campylobacter jejuni into milk has rarely been linked with cases of human infection. We describe the investigations undertaken following the isolation of Campylobacter jejuni from samples of unpasteurized milk prior to retail. Results of epidemiological investigations including typing of Campylobacter jejuni isolates using pyrolysis mass spectrometry, Penner and Lior serotyping, biotyping, phage typing and restriction fragment length polymorphism analysis provided convincing evidence implicating direct milk excretion of Campylobacter jejuni by one asymptomatic dairy cow as the source of the milk contamination and the cause of local cases of human enteritis.

Rapid inter-strain comparison by pyrolysis mass spectrometry in nosocomial infection with Xanthomonas maltophilia

Seventeen strains of Xanthomonas maltophilia and one strain of Pseudomonas cepacia were examined by pyrolysis mass spectrometry (PYMS). The Xanthomonas strains comprised 11 clinical and environmental isolates from a suspected outbreak of colonization and infection on a heart-lung transplant intensive care unit, two strains from patients elsewhere in the same hospital and four strains from a national reference collection. The single isolate of Pseudomonas cepacia was from a sink in the same affected intensive care unit. A series of discriminant analyses performed on the PYMS-derived data showed that, whereas six strains of Xanthomonas from the respiratory tract, blood and ventilatory equipment of one of the affected patients were indistinguishable, all the other isolates were distinct. The results of PYMS rapid inter-strain comparison were in accord with those of an epidemiological investigation which suggested that the episode was due to unauthorized reuse of disposable nebulizers and not to cross-infection between patients. Pyrolysis mass spectrometry with rapid data analysis is a potentially useful technique for the investigation of nosocomial infections due to organisms such as X. maltophilia.

Carbon dioxide laser irradiation of bacterial targets in vitro

Agar targets seeded with Escherichia coli and Staphylococcus aureus in roll tubes simulating the vaginal vault were irradiated with a CO2 laser at various power densities and durations. Viable bacteria were detected in the plume emissions in all instances. Staphylococcus aureus was found to be more resistant to the thermal effects of lasing than E. coli. This suggests that CO2 irradiation of cervical lesions could disseminate viable particles which may be a hazard for patients and operators.

CHARACTERISTICS OF MOTILE CURVED RODS IN VAGINAL SECRETIONS

Motile curved rods seen in vaginal secretions have been isolated on Columbia agar supplemented with 5% human blood and vitamin K. Growth occurred anaerobically and in 5% oxygen but not in more aerobic conditions. There were two distinct groups of these organisms, distinguishable by morphology, biochemical activity and susceptibility to metronidazole. All isolates were sensitive to a wide range of antimicrobial agents, with the exception of nalidixic acid and polymyxin, but one group was resistant to metronidazole. There was little difference between the results of tests of susceptibility to aminoglycosides or to metronidazole performed in anaerobic and microaerophilic conditions. Motile curved rods were isolated from 18 of 80 patients with a clinical diagnosis of non-specific vaginitis, but from only two of 39 without the disease.

Analysis of epidemic and sporadic strains of Listeria monocytogenes by pyrolysis mass spectrometry

Twenty‐six cultures of Listeria monocytogenes, serovar 4b, including 10 from a food‐borne outbreak in Switzerland and sporadic patient and food isolates from both Switzerland and the UK, were compared by pyrolysis mass spectrometry (PMS). This clustered all of the Swiss epidemic isolates with four other isolates indistinguishable from the Swiss strain by phage typing, a phage non‐typable isolate from a Swiss patient not known to be part of the epidemic and two strains (both from Switzerland) of a different phage type. The eight strains excluded from the PMS‐derived cluster were all either known to be unrelated to the epidemic, or of a phage type distinct from the epidemic strain, or both.

Nosocomial infection with Clostridium difficile investigated by pyrolysis mass spectrometry

Fifty-eight isolates of Clostridium difficile from two distinct outbreaks were examined for inter-strain similarity by pyrolysis mass spectrometry (PMS). The first outbreak began on a geriatric acute unit and spread to a long stay geriatric facility. PMS analysis showed that most isolates from both sites were indistinguishable. Isolates obtained in the preceding year from the long stay facility were found to be closely similar to these outbreak isolates. In the second, smaller outbreak on a female medical ward in another general hospital, PMS again showed that a single strain was probably responsible. Representative isolates from these two different outbreaks were shown to be distinct. The ability to compare rapidly large numbers of isolates of C. difficile makes PMS attractive for initial screening in suspected outbreaks, providing important information for outbreak management and allowing conventional typing methods to be concentrated on relevant isolates.

Inter-strain comparison by pyrolysis mass spectrometry in the investigation of Staphylococcus aureus nosocomial infection

Pyrolysis mass spectrometry (PyMS) was used to examine isolates of Staphylococcus aureus from an outbreak of wound infections on a cardiothoracic surgical unit, some of which were thought to have been related to a point-source in the operating theatre. The PyMS results were compared with the results of phage typing. Both methods suggested that a single strain of S. aureus, of phage pattern 29/52/52A/79/80/81, was responsible for some of the wound infections, but PyMS also identified two patients with phage non-typable isolates. Phage typing indicated four staff members as possible carriers of the epidemic strain, but PyMS indicated only two. Epidemiological enquiry confirmed that one of the two members of staff identified by both methods was likely to have been the source of the theatre-based infection. PyMS is a rapid and relatively inexpensive technique for the investigation of nosocomial S. aureus infection and was more discriminatory than phage typing in this instance.

Curie-Point Pyrolysis Mass Spectrometry as a Tool in Clinical Microbiology

Pyrolysis mass spectrometry is a well established analytical tool that has received a considerable boost from the development of low cost, dedicated instruments and sophisticated statistical analyses on personal computers. Further analytical developments, especially in the area of neural networks, are pushing the technology to the forefront of methods for the discrimination and identification of microorganisms and their products. The speed and reproducibility of pyrolysis mass spectrometry and its applicability to a wide range of microorganisms make it an attractive method for epidemiological studies. For inter-strain comparisons, the method is at least as discriminatory as conventional typing systems and usually gives discrimination similar to that of nucleic acid fingerprinting techniques. There has been some success in using neural networks to make identifications across pyrolysis mass spectrometric batches. Further development of methods used to handle data from multiple PyMS analyses can be expected to extend the value of pyrolysis mass spectrometry in clinical microbiology.

Strain differentiation of nosocomial isolates of Pseudomonas aeruginosa by pyrolysis mass spectrometry

Pyrolysis mass spectrometry (PyMS) was used for rapid interstrain comparison of Pseudomonas aeruginosa isolates from a small outbreak of sternal wound infections in a cardiothoracic surgical ward. The PyMS results were compared with those obtained by conventional O-serotyping. All the isolates were phage non-typable. Evidence obtained from PyMS of identity/non-identity between isolates correlated completely with that obtained by O-typing and correctly identified those isolates comprising the epidemic strain, in one instance in advance of O-serotyping. The speed and versatility of PyMS make it an attractive technique for the initial screening of isolates from nosocomial outbreaks of infection with P. aeruginosa and other organisms.