Pol Huedo

Objectives: Estrogen decline during menopause transition is associated with physical and psychological symptoms that impair overall well-being. In a previous randomized trial, a probiotic formula comprising Levilactobacillus brevis KABP052, Lactiplantibacillus plantarum KABP051, and Pediococcus acidilactici KABP021 was found to be associated with a slower decline in estrogen levels. Therefore, we hypothesized that the intake of 1×10 ⁹ colony-forming units/day of this formula could improve menopausal symptoms and quality of life. Methods: We conducted a decentralized, randomized, double-blind, placebo-controlled trial using a real-world approach. Women between 45 and 60 years old who were in perimenopause or postmenopause and had Cervantes Quality of Life score ≥ 38 were enrolled and allocated 2:1 to the probiotic (n=160) or placebo group (n=85) for a 3-month period. Menopausal symptoms and quality of life were evaluated by using the Menopause Rating Scale II, Cervantes and Utian Quality of Life questionnaires. Results: In the full analysis set, Menopause Rating Scale II total score decreased significantly in the probiotic group compared to placebo across the study (global mean difference between groups: −1.00 [0.47], P =0.033), with an improvement in the somato-vegetative domain (−0.54 [0.20], P =0.007). Per-protocol analysis also revealed differences in the psychological domain between groups, favoring the probiotic (−0.51 [0.25], P =0.040). Cervantes scale showed no significant differences between groups, but the change in Utian Quality of Life score was significantly higher in the probiotic group (2.21 [1.07], P =0.040). Conclusion: In this real-world study, the probiotic formula was associated with improvements in menopause-associated symptoms and quality of life, whereas the global Cervantes score appeared to be similar between groups. Future investigations are required to confirm this clinical effect and the underlying mechanisms of action. ClinicalTrials.gov identifier: NCT06604234.

We present the draft genome sequences of Lactiplantibacillus plantarum strains KABP062 and KABP063, two fecal isolates with probiotic potential.

Vaginal dysbiosis is linked to recurrent infections and reproductive complications. Probiotics may restore vaginal microbiota, but there is modest evidence to support vaginal colonization after oral administration. This work aimed to screen a vaginal lactobacilli collection ( n = 45) and assess vaginal colonization of selected candidates. Lactobacillus gasseri CECT 30648 (Lg) and Lactobacillus crispatus CECT 30647 (Lc) showed excellent in vitro attributes, including antagonism against 10 urogenital pathogens, resistance to gastrointestinal tract and vaginal environment—including high concentrations of biogenic amines—and adhesion to vaginal epithelium. Forty-eight healthy women (18–45 years old) were randomly allocated to consume daily a capsule containing Lg (10 ⁹ CFU), a combination of Lg plus Lc (1.5 × 10 ⁹ CFU), or placebo for up to 18 days (between menses). Volunteers collected vaginal samples every 3 days. The primary endpoint was the presence of probiotic strains in vaginal swabs confirmed by strain-specific quantitative PCR. Secondary variables included vaginal microbiota composition and tolerability, among others. Probiotics were well tolerated. Lg but not Lc was detected in 55.9% of participants in active groups (9 in Lg and 10 in Lg + Lc) throughout the study ( P = 0.005 aggregated probiotic groups vs placebo). The sum of relative abundances of non-lactobacilli genera was significantly reduced in the probiotic group after intervention ( P = 0.047 vs day 0), and there was a positive transition toward lactobacilli-dominated community state types ( P = 0.039 vs day 0). Overall, we show that L. gasseri CECT 30648 exerts a broad-spectrum antimicrobial activity and can colonize the vaginal tract of healthy women, thus showing potential to promote vaginal health. The trial was registered in clinicaltrials.gov ( NCT05688397 ). IMPORTANCE: The use of probiotics to promote vaginal health is increasing because vaginal dysbiosis has been linked to numerous gynecological and reproductive complications. While vaginal administration of probiotics using devices or creams has been widely investigated, there is limited evidence supporting vaginal colonization of a probiotic administered orally. It is therefore imperative to perform extensive in vitro characterization to select a vaginal probiotic that can survive the gastrointestinal transit and effectively colonize the vaginal tract of consumers through the oral-gut-vaginal route. We have identified the strain L. gasseri CECT 30648, which shows great probiotic properties, including antagonism against several relevant urogenital pathogens, can colonize the vaginal tract of >55% of participants, and can modulate vaginal microbiota toward a lactobacillus-dominated status in a randomized controlled clinical trial in healthy premenopausal women. These results suggest that oral consumption of L. gasseri CECT 30648 might be effective in promoting vaginal health.

The lactic fermentation of fruit and vegetable juices by well-characterised probiotics remains relatively underexplored. We have investigated the stability and impact of Lactiplantibacillus plantarum KABP051 fermentation on orange, apple, and peach juices by microbiological, physicochemical, and sensory evaluation means. For each fruit juice, three different samples were analysed: original fruit juice without probiotic as blank (B), fruit juice inoculated with 10⁷ CFU/mL of probiotic without fermentation (P), and fruit juice inoculated with 10⁷ CFU/mL of probiotic and fermented at 37 °C for 24 h (PF). P samples displayed good stability throughout the study, and PF samples showed an initial increase in CFUs accompanied by a change in pH, confirming the ability of the probiotic to ferment these juices. After 60 days of refrigeration, PF samples contained >10⁷ CFU/mL. Total phenolic content and antioxidant capacity were equivalent in F, P, and PF. Remarkably, deep metabolomic analyses confirmed malolactic fermentation and revealed the production of several bioactive compounds including the antimicrobial substance phenyllactic acid, the immunomodulatory and anti-fatigue amino acid N-acetyl glutamine, the vitamin B3 form nicotinic acid, the monoterpene (−)-β-pinene, and the neurotransmitter acetylcholine, among others, during probiotic fermentation. Finally, a hedonic analysis involving 51 participants showed that probiotic fermented orange juice is well accepted by panellists, with scores comparable to those of the control juice. Overall, we here show that fruit juices are excellent carriers for the delivery of the probiotic L. plantarum KABP051 and its non-alcoholic fermentation can result in tasty functional fruit juices enriched with health-promoting compounds.

We present the whole-genome sequences of Lactobacillus crispatus CECT30647 and Lactobacillus gasseri CECT30648, two vaginal isolates with probiotic potential.

Probiotics with high bile salt hydrolase (BSH) activity have shown to promote cardiovascular health. However, their mechanism(s) of action remain poorly understood. Here, we performed a pilot exploratory study to investigate effects of a 4-week intervention with escalating doses of a BSH-active formula containing Lactiplantibacillus plantarum strains KABP011, KABP012 and KABP013 on bile acid (BA), lipid profile and lipoprotein function. Healthy overweight individuals were included in this study. The probiotic intake was associated to a progressive decrease of conjugated BAs in serum, due to the reduction of tauro- and glyco-conjugated forms. Plasma levels of Fibroblast Growth Factor-19 (FGF-19) were significantly reduced, and correlated with BA changes. The probiotic induced significant changes in serum lipids, with reduction in non-HDL-cholesterol (non-HDLc) and LDL-cholesterol (LDLc) levels. The largest decrease was evidenced in the subgroup with higher baseline LDL levels (LDLc > 130 mg/dL). Fasting levels of circulating apolipoprotein(Apo) B100 and ApoB48 were significantly reduced. Importantly, the decrease in non-HDLc levels was associated with a significant reduction in small-LDL particles. Functional testing indicated that LDL particles had a significantly lower susceptibility to oxidation whilst HDL particles gained antioxidant capacity after the probiotic intake. The microbiota profile in faeces collected at the end of the study was enriched with members of class Desulfovibrio, a taurine-consuming bacteria, likely because of the increase in free taurine in the gut due to the BSH activity of the probiotic. The intervention with Lactiplantibacillus plantarum strains induces beneficial effects on BA signature and lipoprotein profile. It reduces ApoB and small-LDL levels and LDL susceptibility to oxidation, and increases HDL antioxidant capacity. These metabolic profile changes suggest increased protection against atherosclerotic disease.

To determine the oral colonization capacity of the strains Levilactobacillus brevis KABPTM-052 (CECT 7480) and Lactiplantibacillus plantarum KABPTM-051 (CECT 7481) in healthy subjects. This randomized, double-blinded, placebo-controlled study included 40 volunteers (22 females, 18 males; age range 18-55 years) with healthy gingiva or mild gingivitis, allocated to receiving probiotic chewing gum (n=20) or placebo (n=20) b.i.d for 6 weeks. At baseline and after 6 weeks of treatment, a periodontics specialist collected saliva samples to assess probiotic colonization by qPCR, and analysed dental plaque, gingival index and dental probing pocket depth in Community Periodontal Index (CPI) teeth subset. Protocol was registered as NCT03540498. Treatment compliance was high (99%). Both L. brevis and L. plantarum were detected in the oral microbiota at baseline. After 6 weeks, volunteers receiving probiotic showed a significant increase of both L. brevis (p = 0.017) and L. plantarum (p = 0.004) versus placebo. This effect remained significant after adjusting for gender and gingival index at baseline. In the probiotic group, reduction in plaque index significantly correlated to higher levels of L. brevis (rho = 0.57, p = 0.022) but not of L. plantarum at study endpoint, and the number of subjects with dental plaque was reduced during intervention (7 of 17, p = 0.016). No such effects were observed in the placebo group. No adverse drug reactions were reported. Levilactobacillus brevis KABPTM-052 and Lactiplantibacillus plantarum KABPTM-051 colonize the buccal microbiota of healthy volunteers, and higher colonization by L. brevis positively correlated to reduction in dental plaque. Key words:Probiotic, Levilactobacillus brevis, Lactiplantibacillus plantarum, oral colonization, oral microbiota, dental plaque.

Salmonella enterica serovar Napoli (serovar Napoli) is an emerging cause of human salmonellosis in Northern Italy. No specific reservoirs of serovar Napoli have been identified in Italy, so far. However, the environment, especially surface waters, has been hypothesized as an important source of infection based on the observation that genotypically different clusters of serovar Napoli are detected in different geographical macro-areas. To further support the hypothesis of a spatially-restricted pattern of serovar Napoli diffusion, a spatial segregation of serovar Napoli lineages should be observed also at smaller geographical scale. However, classical genotyping techniques used for Salmonella, such as pulsed-field gel electrophoresis (PFGE), did not possess enough discriminatory power to highlight spatial clustering of serovar Napoli within the macro-areas. To this purpose, we performed phylogeographical analyses based on genome-wide single nucleotide polymorphisms to test whether spatio-temporal evolution patterns of serovar Napoli in Northern Italy could be recognized with high geographical resolution, i.e. at local level. Specifically, we analyzed the local spread of the main PFGE clonal group, responsible for more than 60% of human infections in the study area, that did not show any geographical differentiation by PFGE within Northern Italy, i.e. the macro-area considered in the study. Both discrete and continuous phylogeography highlighted the existence of two main geographically-restricted clades: a Southern clade corresponding to the Po Valley and a Northern clade corresponding to the Pre-Alps area. Furthermore, the phylogeographical analyses suggested that the most probable site of origin of the clone was in an area of the Po Valley at the confluence of the Po and Ticino rivers, one of the most important Italian wetlands. These findings provide further support to the hypothesis that environmental transmission may play an important role in the ecology of serovar Napoli.

Stenotrophomonas maltophilia is an opportunistic Gram-negative pathogen with increasing incidence in clinical settings. The most critical aspect of S. maltophilia is its frequent resistance to a majority of the antibiotics of clinical use. Quorum Sensing (QS) systems coordinate bacterial populations and act as major regulatory mechanisms of pathogenesis in both pure cultures and poly-microbial communities. Disruption of QS systems, a phenomenon known as Quorum Quenching (QQ), represents a new promising paradigm for the design of novel antimicrobial strategies. In this context, we review the main advances in the field of QS in S. maltophilia by paying special attention to Diffusible Signal Factor (DSF) signaling, Acyl Homoserine Lactone (AHL) responses and the controversial Ax21 system. Advances in the DSF system include regulatory aspects of DSF synthesis and perception by both rpf-1 and rpf-2 variant systems, as well as their reciprocal communication. Interaction via DSF of S. maltophilia with unrelated organisms including bacteria, yeast and plants is also considered. Finally, an overview of the different QQ mechanisms involving S. maltophilia as quencher and as object of quenching is presented, revealing the potential of this species for use in QQ applications. This review provides a comprehensive snapshot of the interconnected QS network that S. maltophilia uses to sense and respond to its surrounding biotic or abiotic environment. Understanding such cooperative and competitive communication mechanisms is essential for the design of effective anti QS strategies.

Molecular subtyping and enhanced surveillance in Lombardy region identified a cluster of possibly related listeriosis cases from 2006 to 2010. This cluster grouped 31 isolates that belonged to serotype 1/2a and Sequence Type 38 (ST38) as defined by Multilocus Sequence Typing (MLST). Our study expanded the previous investigation to include cases from 2011 to 2014 and used Multi-Virulence-Locus Sequence Typing (MVLST) on all ST38 isolates to better understand their epidemiology and possibly identify a common source outbreak. Out of 306 L. monocytogenes clinical isolates collected, 43 (14.1%) belonged to ST38 with cases occurring in nine out of twelve Lombardy provinces. The ST38 isolates were split by MVLST into two Virulence Types (VTs): VT80 (n = 12) and VT104 (n = 31). VT104 cases were concentrated between 2009 and 2011 in two provinces, Bergamo and Milan. An epidemiologic investigation was performed and in one case, a matching VT104 isolate was retrieved from a soft cheese sample from a patient’s refrigerator. Our findings revealed a major listeriosis outbreak in Northern Italy linked to soft cheese in 2009–2011, which went undetected by local health authorities. Our study shows that integrating subtyping methods with conventional epidemiology can help identify the source of L. monocytogenes outbreak clones.