Genetic characterization and antibiotic resistance of Campylobacter jejuni isolated from meats, water, and humans in Sweden.

The incidence of Campylobacter jejuni has increased during the last decade, and today it is the leading cause of bacterial enteritis in most developed countries. Still, there is a lack of knowledge about infection routes and to what extent identified sources are responsible for spreading the bacterium to humans. The major objective of this work was to explore the genetic similarity between C. jejuni isolated from different sources. C. jejuni isolated from patients (n = 95), five types of meat (n = 71), and raw water (n = 11) during the year 2000 were subtyped by pulsed-field gel electrophoresis (PFGE). The pulsotypes obtained after digestion with SmaI revealed not only that C. jejuni is genetically diverse but also that specific pulsotypes occur frequently. Five clusters comprising 88 of the 162 SmaI-digested isolates were obtained. After digestion with KpnI most isolates in four of the five clusters were still indistinguishable, while the fifth cluster was strongly dissolved. The clusters comprised high frequencies of human and meat isolates, while only one of nine water isolates belonged to a cluster. The largest cluster comprised 21 human isolates, one raw water isolate, and seven chicken meat isolates, originating from at least six different broiler flocks. Low frequencies of antibiotic resistance were revealed when the meat and water isolates were tested for sensitivity to six antibiotics. Interestingly, the five isolates resistant to quinolones displayed similar or identical pulsotypes. The results showed that PFGE has proved useful in identifying clones and will be used in future work focusing on identification and eradication of the major reservoirs for common clones.

Quantitative risk assessment for Listeria monocytogenes in smoked or gravad salmon and rainbow trout in Sweden.

The objective of the present work was to develop a quantitative risk assessment model in which the exposure and risk of acquiring listeriosis from consumption of packaged smoked or gravad salmon and rainbow trout were estimated. An Excel spreadsheet model was constructed in which variables were represented by distributions based on surveys of L. monocytogenes in these food products, and on demographic and consumption data. Growth or inactivation was not included in the model. The model was run through Monte Carlo simulations using the @Risk software (Palisade Corporation). The probability of illness per serving was calculated using two dose-response models from the literature. The first was an exponential model in which the species specific constant R, that helps define the dose-response curve, previously has been estimated to be 1.18 x 10(-10) based on German data (GR). In this study, R was estimated to 5.6 x 10(-10) based on Swedish data. The second model was a flexible Weibull-Gamma model (WG), with different coefficients for high- and low-risk groups. The exponential model (GR), although conservative and generally overestimating the risk, still predicted a lower probability of illness than the WG-model. The estimated mean risk per serving was 2.8 x 10(-5) (GR, high-risk group), 2.0 x 10(-3) (WG, low-risk group) and 0.016 (WG, high-risk group), respectively. The average number of reported listeriosis cases in Sweden is 37 per year. In comparison, the mean number of annual cases predicted by the risk assessment model was 168 (range 47 to 2800, GR, high-risk group), and 95 000 (range 34 000 to 1.6 x 10(6), WG high-risk group), respectively. If 1 to 10% (uniform distribution) of strains, instead of all, were considered virulent, the mean number of predicted cases would decrease to nine (GR) and 5200 (WG), respectively. The mean annual cumulative individual risk in the high-risk group based on a monthly exposure was estimated to be 4.0 x 10(-4) (range 8.0 x 10(-8) to 5.4 x 10(-3), GR). This risk increased to 1.5 x 10(-3) (range 1.7 x 10(-5) to 9.2 x 10(-3), GR) based on a weekly exposure. The risk assessment model was most sensitive to the input distribution describing the level of contamination and to a lesser degree on the prevalence of L. monocytogenes, the proportion of virulent strains, and serving sizes. A lack of data on the prevalence and concentration of L. monocytogenes in these products, dose-response data and quantitative information on the proportion of virulent strains were identified.

Cloning of the immunity repressor determinant of bacteriophage P2 in the pBR322 plasmid.

Through in vitro recombination of DNA restriction fragments, we have constructed a plasmid, which expressed in vivo the immunity repressor gene (C) of bacteriophage P2. A bacterial strain carrying such a plasmid showed a high level of P2 specific immunity. It was lysogenized normally by an infecting P2, but the frequency of spontaneous phage production was reduced about 10(4) fold as compared to a normal P2 lysogen. Satellite phage P4, known to derepress P2 lysogens, was unable to derepress the plasmid-carrying lysogenic strain so to allow growth of coinfecting P2. Phage P4 multiplied on the plasmid-carrying, P2-lysogenic strain, but due to a prolonged latent period failed to form plaques on this strain.

A restriction endonuclease cleavage map of bacteriophage P2.

A restriction endonuclease cleavage map of phage P2 was constructed. The enzymes used and, within parenthesis, the number of their cleavage sites on the P2 lg cc DNA molecule were: AvaI(3), BalI(1), BAMI(3), BglII(3), HaeIII (more than 40; only three were mapped), HindIII(0), HpaI(10), KpnI(3), PstI(3), SalI(2) and SmaI(2). The EcoRI cleavage sites (3), as determined earlier, were used as reference points for this study. The DNAs of a variety of P2 mutants carrying chromosomal aberrations (dell, del2, del3, del6, vir22, vir37(2), vir79 and vir94) were also similarly examined.