Novel ways to use sensor data to improve mastitis management.

Current sensor systems are used to detect cows with clinical mastitis. Although, the systems perform well enough to not negatively affect the adoption of automatic milking systems, the performance is far from perfect. An important advantage of sensor systems is the availability of multiple measurements per day. By clearly defining the need for detection of subclinical mastitis (SCM) and clinical mastitis (CM) from the farmers' management perspective, detection and management of SCM and CM may be improved. Sensor systems may also be used for other aspects of mastitis management. In this paper we have defined 4 mastitis situations that could be managed with the support of sensor systems. Because of differences in the associated management and the epidemiology of these specific mastitis situations, the required demands for performance of the sensor systems do differ. The 4 defined mastitis situations with the requirements of performance are the following: (1) Cows with severe CM needing immediate attention. Sensor systems should have a very high sensitivity (>95% and preferably close to 100%) and specificity (>99%) within a narrow time window (maximum 12 h) to ensure that close to all cows with true cases of severe CM are detected quickly. Although never studied, it is expected that because of the effects of severe CM, such a high detection performance is feasible. (2) Cows with mastitis that do not need immediate attention. Although these cows have a risk of progressing into severe CM or chronic mastitis, they should get the chance to cure spontaneously under close monitoring. Sensor alerts should have a reasonable sensitivity (>80%) and a high specificity (>99.5%). The time window may be around 7 d. (3) Cows needing attention at drying off. For selective dry cow treatment, the absence or presence of an intramammary infection at dry-off needs to be known. To avoid both false-positive and false-negative alerts, sensitivity and specificity can be equally high (>95%). (4) Herd-level udder health. By combining sensor readings from all cows in the herd, novel herd-level key performance indicators can be developed to monitor udder health status and development over time and raise alerts at significant deviances from predefined thresholds; sensitivity should be reasonably high, >80%, and because of the costs for further analysis of false-positive alerts, the specificity should be >99%. The development and validation of sensor-based algorithms specifically for these 4 mastitis situations will encourage situation-specific farmer interventions and operational udder health management.

Effects of glucose and oxygen on arginine metabolism by coagulase-negative staphylococci.

Coagulase-negative staphylococci (CNS) are not only part of the desirable microbiota of fermented meat products but also commonly inhabit skin and flesh wounds. Their proliferation depends on the versatility to use energy sources and the adaptation to fluctuating environmental parameters. In this study, the conversion of the amino acid arginine by two strains with arginine deiminase (ADI) activity (Staphylococcus carnosus 833 and S. pasteuri αs3-13) and a strain with nitric oxide synthase (NOS) activity (S. haemolyticus G110) was modelled as a function of glucose and oxygen availability. Both factors moderately inhibited the ADI-based conversion kinetics, never leading to full repression. However, for NOS-driven conversion of arginine by S. haemolyticus G110, oxygen was an absolute requirement. When changing from microaerobic conditions to aerobiosis, a switch from homolactic fermentation to a combined formation of lactic acid, acetic acid, and acetoin was found in all cases, after which lactic acid and acetic acid were used as substrates. The kinetic model proposed provided a suitable description of the data of glucose and arginine co-metabolism as a function of oxygen levels and may serve as a tool to further analyse the behaviour of staphylococci in different ecosystems or when applying specific food processing conditions.

Coagulase-negative Staphylococci favor conversion of arginine into ornithine despite a widespread genetic potential for nitric oxide synthase activity.

Within ecosystems that are poor in carbohydrates, alternative substrates such as arginine may be of importance to coagulase-negative staphylococci (CNS). However, the versatility of arginine conversion in CNS remains largely uncharted. Therefore, a set of 86 strains belonging to 17 CNS species was screened for arginine deiminase (ADI), arginase, and nitric oxide synthase (NOS) activities, in view of their ecological relevance. In fermented meats, for instance, ADI could improve bacterial competitiveness, whereas NOS may serve as an alternative nitrosomyoglobin generator to nitrate and nitrite curing. About 80% of the strains were able to convert arginine, but considerable inter- and intraspecies heterogeneity regarding the extent and mechanism of conversion was found. Overall, ADI was the most commonly employed pathway, resulting in mixtures of ornithine and small amounts of citrulline. Under aerobic conditions, which are more relevant for skin-associated CNS communities, several strains shifted toward arginase activity, leading to the production of ornithine and urea. The obtained data indeed suggest that arginase occurs relatively more in CNS isolates from a dairy environment, whereas ADI seems to be more abundant in strains from a fermented meat background. With some exceptions, a reasonable match between phenotypic ADI and arginase activity and the presence of the encoding genes (arcA and arg) was found. With respect to the NOS pathway, however, only one strain (Staphylococcus haemolyticus G110) displayed phenotypic NOS-like activity under aerobic conditions, despite a wide prevalence of the NOS-encoding gene (nos) among CNS. Hence, the group of CNS displays a strain- and condition-dependent toolbox of arginine-converting mechanisms with potential implications for competitiveness and functionality.

Exploring the metabolic heterogeneity of coagulase-negative staphylococci to improve the quality and safety of fermented meats: a review.

The production of fermented meats, such as fermented sausage, relies on the metabolic activities of lactic acid bacteria and catalase-positive cocci, in particular the group of coagulase-negative staphylococci (CNS). Conventional use of CNS as meat starter cultures usually leads to an appropriate cured colour development based on their nitrate reductase activity, whereas their catalase activity reduces oxidative damage. In addition, CNS metabolism contributes to flavour, although the precise effects are difficult to estimate. There are reasons to believe that these basic technological features of CNS can be further enlarged by exploring their full metabolic potential. Non-negligible differences in metabolism among and within different species of CNS indicate that a rational selection of strains may lead to the development of novel starter cultures with enhanced functionality. Firstly, the use of CNS strains with a superior ability to use alternative energy sources, such as nucleosides or arginine, may improve culture competitiveness and survival. Secondly, cured colour generation could be optimised to lower the amounts of curing salts needed, either by selecting for efficient nitrate-reducing CNS strains or by exploring the potential alternative based on nitric oxide synthase activity. Thirdly, CNS with specific aroma-producing abilities may help to accentuate specific flavours, whereby the selection of wild-type strains from artisan-type fermented sausages seems attractive in the framework of innovation-through-tradition. Finally, bacteriocin-producing CNS strains may offer solutions for bioprotection towards meat pathogens such as Clostridium botulinum and Staphylococcus aureus. Overall, making use of the metabolic inter- and intraspecies heterogeneity of CNS is promising for the elaboration of healthier, tastier, and safer fermented meats. Yet, the proposed strategies are sometimes still overly theoretical and speculative, requiring further proof-of-principle.

Process-driven bacterial community dynamics are key to cured meat colour formation by coagulase-negative staphylococci via nitrate reductase or nitric oxide synthase activities.

The cured colour of European raw fermented meats is usually achieved by nitrate-into-nitrite reduction by coagulase-negative staphylococci (CNS), subsequently generating nitric oxide to form the relatively stable nitrosomyoglobin pigment. The present study aimed at comparing this classical curing procedure, based on nitrate reductase activity, with a potential alternative colour formation mechanism, based on nitric oxide synthase (NOS) activity, under different acidification profiles. To this end, meat models with and without added nitrate were fermented with cultures of an acidifying strain (Lactobacillus sakei CTC 494) and either a nitrate-reducing Staphylococcus carnosus strain or a rare NOS-positive CNS strain (Staphylococcus haemolyticus G110), or by relying on the background microbiota. Satisfactory colour was obtained in the models prepared with added nitrate and S. carnosus. In the presence of nitrate but absence of added CNS, however, cured colour was only obtained when L. sakei CTC 494 was also omitted. This was ascribed to the pH dependency of the emerging CNS background microbiota, selecting for nitrate-reducing Staphylococcus equorum strains at mild acidification conditions but for Staphylococcus saprophyticus strains with poor colour formation capability when the pH decrease was more rapid. This reliance of colour formation on the composition of the background microbiota was further explored by a side experiment, demonstrating the heterogeneity in nitrate reduction of a set of 88 CNS strains from different species. Finally, in all batches prepared with S. haemolyticus G110, colour generation failed as the strain was systematically outcompeted by the background microbiota, even when imposing milder acidification profiles. Thus, when aiming at colour formation through CNS metabolism, technological processing can severely interfere with the composition and functionality of the meat-associated CNS communities, for both nitrate reductase and NOS activities. Several major bottlenecks, among which the rareness of phenotypic NOS activity in meat-compatible CNS, need to be considered, which is seriously questioning the relevance of this pathway in fermented meats.