Initial Transfer Event Research Articles

Quantifying the uncertainty of transfer of Cronobacter spp. between fomites and floors and touch points in dairy processing plants

Cronobacter spp. are opportunistic pathogens that must be controlled in infant powder manufacturing plants. This study evaluated the spread of Cronobacter cells via contact surfaces within a dairy manufacturing environment. Transfer rates of Cronobacter spp. were determined from vectors for transmission including moveable fomites (e.g. trolley wheels and boots) and gloved hands to various types of recipient surfaces (stainless steel, linoleum and resin-coated concrete) typical for dairy manufacturing environments. Overall, with a starting inoculum of 106 CFU/mL, approximately 104 CFU/mL Cronobacter cells were transferred from each fomite onto each recipient surface during the initial transfer event. Gloved hands transferred the highest number of Cronobacter cells, followed by polyvinylchloride boots and then polyurethane trolley wheels. We demonstrate, using a combination of experimental data and uncertainty analysis, that if a movable fomite (boots or trolley wheels), or gloves became contaminated, Cronobacter could be spread over a wide area within a manufacturing plant. To the authors’ knowledge, this is the first quantitative estimation of the spread of Cronobacter within a dairy manufacturing plant, that can also be practically applied as a tool for providing information in making risk management decisions. In particular, the estimation of spread suggests areas for cleaning and sanitation within a dairy manufacturing environment during a contamination event.

Transfer during unlubricated sliding wear of selected metal systems

Results of pin-on-disk sliding tests in vacuum, with copper, CuNi, nickel and molybdenum sliding against iron, indicate that initial transfer events involve discrete fragments. We propose that the process involved in the initial transfer events is a consequence of local shear instabilities which develop at large plastic strains. In copper samples the initial transfer elements are lamellar, with layer thickness equal to the cell thickness in the highly deformed base material adjacent to the sliding interface. Prolonged sliding gives rise to the formation of transfer layers or patches on the specimen surface. These layers or patches are composed of finely mixed material derived from the two sliding counterparts as well as scattered pieces of more recently transferred fragments. Typical wear debris particles are generated from the transfer layers or patches. Transfer tendencies for different materials combinations can be predicted from an adhesion point of view if geometrical effects are properly considered. Combining theoretical predictions of transfer tendencies and experimental observations of transfer and geometrical effects, one finds that for a pin-and-disk system the disk should be made of the material having the higher cohesive strength.

Measurement of transcribed human X-chromosomal DNA sequences transferred to rodent cells by chromosome-mediated gene transfer.

Transfer of genetic information can be effected by incubation of cultured eucaryotic cells with isolated metaphase chromosomes. In most cases, a resulting transformed cell contains only a fragment of a donor chromosome. The amount of transferred donor DNA has been quantified in 11 independent mouse A9 transformants by nucleic acid hybridization analysis. Each transformant had been selected for hprt (hypoxanthine phosphoribosyltransferase; EC 2.4.2.8) transfer and contained part of the human X chromosome. A labeled probe of transcribed human X-chromosomal DNA was prepared by hybridization of nick-translated unique-sequence human DNA with whole cellular RNA from a human-mouse hybrid cell line, A9/HRBC2-A, containing a single human chromosome., X. The amount of human X-chromosomal DNA in the transformants was quantitated by comparing the hybridization of this probe with transformant and A9/HRBC2-A DNAs. Two unstable transformants which had a microscopically detectable donor chromosome fragment contained 15% of the human X-chromosomal single-copy DNA. Four other unstable transformants contained 4 to 7% of human X-chromosomal DNA sequences. The transferred DNA was below the level of detection in three other unstable and in all three stable transformants. We conclude that the initial transfer event can introduce a substantial amount of genetic information but only smaller amounts of DNA are stably incorporated by integration.

Measurement of transcribed human X-chromosomal DNA sequences transferred to rodent cells by chromosome-mediated gene transfer.

Transfer of genetic information can be effected by incubation of cultured eucaryotic cells with isolated metaphase chromosomes. In most cases, a resulting transformed cell contains only a fragment of a donor chromosome. The amount of transferred donor DNA has been quantified in 11 independent mouse A9 transformants by nucleic acid hybridization analysis. Each transformant had been selected for hprt (hypoxanthine phosphoribosyltransferase; EC 2.4.2.8) transfer and contained part of the human X chromosome. A labeled probe of transcribed human X-chromosomal DNA was prepared by hybridization of nick-translated unique-sequence human DNA with whole cellular RNA from a human-mouse hybrid cell line, A9/HRBC2-A, containing a single human chromosome., X. The amount of human X-chromosomal DNA in the transformants was quantitated by comparing the hybridization of this probe with transformant and A9/HRBC2-A DNAs. Two unstable transformants which had a microscopically detectable donor chromosome fragment contained 15% of the human X-chromosomal single-copy DNA. Four other unstable transformants contained 4 to 7% of human X-chromosomal DNA sequences. The transferred DNA was below the level of detection in three other unstable and in all three stable transformants. We conclude that the initial transfer event can introduce a substantial amount of genetic information but only smaller amounts of DNA are stably incorporated by integration.

Factors influencing the establishment and spread of R-plasmids in an experimental model of urinary tract infection.

An experimental model simulating infection of the urinary tract was used to investigate factors influencing the transfer of plasmid-mediated drug resistance in the human bladder in the absence of antibiotic therapy. When a small number of cells carrying plasmid-mediated drug resistance was added to an established population of sensitive cells, it was found that varying the physical conditions of the bladder model had no significant effect on the spread of the plasmid and, under all conditions used, exponential spread of the plasmid through the originally sensitive population occurred. When the initial donor cells were killed shortly after addition to the model, exponential transfer of resistance amongst the established sensitive population still occurred. Thus, even in the absence of antibiotic therapy, only a small number of initial transfer events was required for a fully resistant population to be developed. This was true of both actively multiplying and early stationary phase cultures. An initially greater proportion of resistant cells, or the use of an antimicrobial agent to which some of the cells are resistant, would ensure that this process occurred more rapidly.