Chronic release of tailless phage particles from Lactococcus lactis

Lactococcus lactis strains residing in the microbial community of a complex dairy starter culture named “Ur” are hosts to prophages belonging to the family Siphoviridae . L. lactis strains (TIFN1 to TIFN7) showed detectable spontaneous phage production and release (10 9 -10 10 phage particles/mL) and up to 10-fold increases upon prophage induction, while in both cases we observed no obvious cell lysis, typically described for the lytic life cycle of Siphoviridae phages. Intrigued by this phenomenon, we investigated the host-phage interaction using strain TIFN1 (harboring prophage proPhi1) as a representative. We confirmed that during the massive phage release, all bacterial cells remain viable. Further, by monitoring phage replication in vivo , using a green fluorescence protein reporter combined with flow cytometry, we demonstrated that the majority of the bacterial population (over 80%) is actively producing phage particles when induced with mitomycin C. The released tailless phage particles were found to be engulfed in lipid membranes, as evidenced by electron microscopy and lipid staining combined with chemical lipid analysis. Based on the collective observations, we propose a model of phage-host interaction in L. lactis TIFN1, where the phage particles are engulfed in membranes upon release, thereby leaving the producing host intact. Moreover, we discuss possible mechanisms of chronic, or non-lytic release of LAB Siphoviridae phages and its impact on the bacterial host. Importance: In complex microbial consortia such as fermentation starters, bacteriophages can alter the dynamics and diversity of microbial communities. Bacteriophages infecting Lactococcus lactis are mostly studied for their detrimental impact on industrial dairy fermentation processes. In the submitted article, we describe a novel way of phage-bacteria interaction in a L. lactis strain isolated from a complex dairy starter culture: when the prophages harbored in the L. lactis genome are activated, the phage particles are engulfed in lipid membranes upon release, leaving the producing host intact. Findings from this study provide additional insights into the diverse manners of phage-bacteria interactions and co-evolution, which is essential for understanding the population dynamics in complex microbial communities like fermentation starters.

Lacticaseibacillus rhamnosus Impedes Growth of Listeria spp. in Cottage Cheese through Manganese Limitation

Acidification and nutrient depletion by dairy starter cultures is often sufficient to prevent outgrowth of pathogens during post-processing of cultured dairy products. In the case of cottage cheese, however, the addition of cream dressing to the curd and subsequent cooling procedures can create environments that may be hospitable for the growth of Listeria monocytogenes. We report on a non-bacterio-cinogenic Lacticaseibacillus rhamnosus strain that severely limits the growth potential of L. monocytogenes in creamed cottage cheese. The main mechanism underlying Listeria spp. inhibition was found to be caused by depletion of manganese (Mn), thus through competitive exclusion of a trace element essential for the growth of many microorganisms. Growth of Streptococcus thermophilus and Lactococcus lactis that constitute the starter culture, on the other hand, were not influenced by reduced Mn levels. Addition of L. rhamnosus with Mn-based bioprotective properties during cottage cheese production therefore offers a solution to inhibit undesired bacteria in a bacteriocin-independent fashion.

BENEFICIAL EFFECTS OF PROBIOTICS BASED THERAPY FOR IMMUNE HOMEOSTASIS

Several approaches have been used to demonstrate that signals derived from gut microbiota are critical for the development of the immune system. Probiotics are live microorganisms that have beneficial effects on host health, including extended lifespan, when they are administered or present in adequate quantities. However, the mechanisms by which probiotics stimulate host longevity remain unclear and very poorly understood. Probiotic supplements are needed to determine the effectiveness as a non-chemical approach to promote health and welfare. Our studies focus on the assessment of the molecular impact of probiotic administration involved in homeostasis and immunity. Shrimps received the recommended doses of microbial pack containing Bacillus species andRhodococcus species. The fermentation was carried at 37°C for 72 hours under microaerophilic condition. The parameters like pH, microbial load, acidity and concentration of reducing sugar had been measured.This probiotic pack when used either alone or in combination with traditional dairy starter, significantly improved the nutritional properties and the shelf life of the product. The present work will be valuable to elaborate novel functional food based on these original probiotic properties.

Dairy lactococcal and streptococcal phage–host interactions: an industrial perspective in an evolving phage landscape

ABSTRACT Almost a century has elapsed since the discovery of bacteriophages (phages), and 85 years have passed since the emergence of evidence that phages can infect starter cultures, thereby impacting dairy fermentations. Soon afterward, research efforts were undertaken to investigate phage interactions regarding starter strains. Investigations into phage biology and morphology and phage–host relationships have been aimed at mitigating the negative impact phages have on the fermented dairy industry. From the viewpoint of a supplier of dairy starter cultures, this review examines the composition of an industrial phage collection, providing insight into the development of starter strains and cultures and the evolution of phages in the industry. Research advances in the diversity of phages and structural bases for phage–host recognition and an overview of the perpetual arms race between phage virulence and host defense are presented, with a perspective toward the development of improved phage-resistant starter culture systems.

Complete Genome Sequence of Lactococcus lactis subsp. cremoris 3107, Host for the Model Lactococcal P335 Bacteriophage TP901-1

The complete genome sequence of Lactococcus lactis subsp. cremoris 3107, a dairy starter strain and a host for the model lactococcal P335 bacteriophage TP901-1, is reported here.

High-throughput pH monitoring method for application in dairy fermentations

Optimization of dairy fermentation processes often requires multiplexed pH measurements over several hours. The method developed here measures up to 90 samples simultaneously, where traditional electrode-based methods require a lot more time for handing the same number of samples. Moreover, the new method employs commonly used materials and can be used with a wider range of fluorescence readers than commercial 96-well plates with optical pH sensors. For this application, a milk-like transparent medium is developed that shows acidification properties with dairy starters that are similar to milk. Combination of this milk-like medium and 3 fluorescent indicators allow precise measurements of pH in a range of 4·0–7·0. The new method showed much higher throughput compared to the benchmark electrode systems while being as accurate, as shown by successful application for a comparison of various dairy starter cultures and for optimizing the inoculation rate.