Genomic Characterization of Dairy Associated Leuconostoc Species and Diversity of Leuconostocs in Undefined Mixed Mesophilic Starter Cultures

Undefined mesophilic mixed (DL-type) starter cultures are composed of predominantly Lactococcus lactis subspecies and 1–10% Leuconostoc spp. The composition of the Leuconostoc population in the starter culture ultimately affects the characteristics and the quality of the final product. The scientific basis for the taxonomy of dairy relevant leuconostocs can be traced back 50 years, and no documentation on the genomic diversity of leuconostocs in starter cultures exists. We present data on the Leuconostoc population in five DL-type starter cultures commonly used by the dairy industry. The analyses were performed using traditional cultivation methods, and further augmented by next-generation DNA sequencing methods. Bacterial counts for starter cultures cultivated on two different media, MRS and MPCA, revealed large differences in the relative abundance of leuconostocs. Most of the leuconostocs in two of the starter cultures were unable to grow on MRS, emphasizing the limitations of culture-based methods and the importance of careful media selection or use of culture independent methods. Pan-genomic analysis of 59 Leuconostoc genomes enabled differentiation into twelve robust lineages. The genomic analyses show that the dairy-associated leuconostocs are highly adapted to their environment, characterized by the acquisition of genotype traits, such as the ability to metabolize citrate. In particular, Leuconostoc mesenteroides subsp. cremoris display telltale signs of a degenerative evolution, likely resulting from a long period of growth in milk in association with lactococci. Great differences in the metabolic potential between Leuconostoc species and subspecies were revealed. Using targeted amplicon sequencing, the composition of the Leuconostoc population in the five commercial starter cultures was shown to be significantly different. Three of the cultures were dominated by Ln. mesenteroides subspecies cremoris. Leuconostoc pseudomesenteroides dominated in two of the cultures while Leuconostoc lactis, reported to be a major constituent in fermented dairy products, was only present in low amounts in one of the cultures. This is the first in-depth study of Leuconostoc genomics and diversity in dairy starter cultures. The results and the techniques presented may be of great value for the dairy industry

Upper limb activity in myoelectric prosthesis users is biased towards the intact limb and appears unrelated to goal-directed task performance

Studies of the effectiveness of prosthetic hands involve assessing user performance on functional tasks in the lab/clinic, sometimes combined with self-report of real-world use. In this paper we compare real-world upper limb activity between a group of 20 myoelectric prosthesis users and 20 anatomically intact adults. Activity was measured from wrist-worn accelerometers over a 7-day period. The temporal patterns in upper limb activity are presented and the balance of activity between the two limbs quantified. We also evaluated the prosthesis users’ performance on a goal-directed task, characterised using measures including task success rate, completion time, gaze behaviour patterns, and kinematics (e.g. variability and patterns in hand aperture). Prosthesis users were heavily reliant on their intact limb during everyday life, in contrast to anatomically intact adults who demonstrated similar reliance on both upper limbs. There was no significant correlation between the amount of time a prosthesis was worn and reliance on the intact limb, and there was no significant correlation between either of these measures and any of the assessed kinematic and gaze-related measures of performance. We found participants who had been prescribed a prosthesis for longer to demonstrate more symmetry in their overall upper limb activity, although this was not reflected in the symmetry of unilateral limb use. With the exception of previously published case studies, this is the first report of real world upper limb activity in myoelectric prosthesis users and confirms the widely held belief that users are heavily reliant on their intact limb

EMG Biofeedback for online predictive control of grasping force in a myoelectric prosthesis

Background Active hand prostheses controlled using electromyography (EMG) signals have been used for decades to restore the grasping function, lost after an amputation. Although myocontrol is a simple and intuitive interface, it is also imprecise due to the stochastic nature of the EMG recorded using surface electrodes. Furthermore, the sensory feedback from the prosthesis to the user is still missing. In this study, we present a novel concept to close the loop in myoelectric prostheses. In addition to conveying the grasping force (system output), we provided to the user the online information about the system input (EMG biofeedback). Methods As a proof-of-concept, the EMG biofeedback was transmitted in the current study using a visual interface (ideal condition). Ten able-bodied subjects and two amputees controlled a state-of-the-art myoelectric prosthesis in routine grasping and force steering tasks using EMG and force feedback (novel approach) and force feedback only (classic approach). The outcome measures were the variability of the generated forces and absolute deviation from the target levels in the routine grasping task, and the root mean square tracking error and the number of sudden drops in the force steering task. Results During the routine grasping, the novel method when used by able-bodied subjects decreased twofold the force dispersion as well as absolute deviations from the target force levels, and also resulted in a more accurate and stable tracking of the reference force profiles during the force steering. Furthermore, the force variability during routine grasping did not increase for the higher target forces with EMG biofeedback. The trend was similar in the two amputees. Conclusions The study demonstrated that the subjects, including the two experienced users of a myoelectric prosthesis, were able to exploit the online EMG biofeedback to observe and modulate the myoelectric signals, generating thereby more consistent commands. This allowed them to control the force predictively (routine grasping) and with a finer resolution (force steering). The future step will be to implement this promising and simple approach using an electrotactile interface. A prosthesis with a reliable response, following faithfully user intentions, would improve the utility during daily-life use and also facilitate the embodiment of the assistive system.peerReviewe