Clear aligners have gained popularity in orthodontics due to their aesthetics, comfort, and removability; however, their prolonged intraoral wear and frequent removal–reinsertion cycles create favorable conditions for microbial colonization. This in vitro study evaluated the efficacy of seven commercially available mouthwash formulations in inhibiting biofilms of Streptococcus mutans, Streptococcus oralis, and Candida albicans formed on four different clear aligner materials. Standardized aligner fragments were incubated for 24 h with microbial suspensions to allow biofilm formation, treated for 1 min with one of the mouthwashes, and then assessed for residual viability through spectrophotometric optical density measurements after a further 24 h incubation. Biofilm inhibition varied according to both mouthwash composition and aligner material. The chlorhexidine-based rinse (MW-D) consistently showed the highest inhibition across microorganisms, while the fluoride–cetylpyridinium chloride rinse (MW-B) performed strongly for S. oralis and C. albicans. An essential oil-based formulation with xylitol (MW-G) showed notable antifungal activity against C. albicans. Monolayer polyurethane aligners generally achieved higher inhibition rates than multilayer or copolyester-based materials. These findings indicate that antimicrobial efficacy on aligners depends on both mouthwash type and material, supporting a tailored approach to biofilm management in clear aligner therapy to reduce the risk of caries, periodontal disease, and candidiasis.

The ever-growing global demand for animal protein forces the aquaculture industry to expand at a pace which imposes significant challenges in maintaining sustainable practices. This study aimed to investigate the efficacy of an organic acid mixture (Aq) in mitigating Lactococcus garvieae (L. garvieae) virulence through its effects on bacterial virulence (EPS production, biofilm, and haemolytic factors) and host pathogenicity, including its adherence to CHSE-214 cells, haemolysis, and proinflammatory responses. Our findings reveal that Aq significantly inhibits L. garvieae growth at a 0.125% concentration, suppresses EPS-related gene expression, reduces biofilm formation, and reduces cytotoxicity in fish epithelial cells (CHSE-214). Moreover, Aq decreased haemolysing gene expression (hly1, hly2 and hly3) and attenuated red blood cell haemolysis, a hallmark of L. garvieae pathogenicity. Lastly, Aq was demonstrated to induce modulation in the host immune responses, lowering IL-1β and IL-8 expression, which are critical mediators of inflammation and pathogen recruitment. Therefore, we conclude that the main mechanism of action of Aq involves inhibiting L. garvieae adhesion to epithelial cells, reducing EPS production, and downregulating key virulence-associated genes (e.g., hly1, hly2, and hly3). After preventing L. garvieae adherence and suppressing proinflammatory cytokine expression (IL-1β and IL-8), Aq disrupts the pathogen’s ability to breach epithelial barriers and induce red blood cell lysis, thereby mitigating its virulence and pathogenicity. Our results emphasised the potential of Aq as an alternative non-antibiotic intervention for controlling piscine lactococcosis, advancing our understanding of L. garvieae pathogenesis and providing the foundation for the future integration of environmentally friendly antimicrobials into aquaculture disease management.

Some Vibrio parahaemolyticus strains cause translucent post-larvae disease (Vp_TPD), leading to significant economic losses in shrimp farming. We aimed to identify whether a mixture of natural antimicrobials, AuraAqua (Aq), can protect white-leg shrimp (Penaeus vannamei) against the lethal effects of Vp_TPD and to understand its biological mode of action. Herein, we demonstrate that Aq, an antimicrobial mixture composed of a blend of organic acids, citrus, and olive extracts, suppressed Vp_TPD virulence at sub-inhibitory concentrations and conferred robust protection to shrimp. The minimum inhibitory and bactericidal concentrations against the Vp_TPD isolate were at 0.05% and 0.2%, respectively. At 0.05–0.1%, Aq reduced bacterial growth and downregulated six major virulence genes (vhvp-1, vhvp-2, vhvp-3, pirA_Vp, pirB_Vp, pirAB_Vp), while leaving metabolic ldh expression unaltered. Parallel in vitro assays revealed diminished adhesion of Vp_TPD to primary shrimp gut and hepatopancreas epithelial cells and a ≈50% reduction in infection-induced extracellular H₂O₂, indicating an antioxidant effect. The treatment also triggered a time-dependent surge in extracellular alkaline phosphatase (ALP) activity, consistent with membrane permeabilization. In vivo, a challenge of post-larvae with 10⁴ CFU/mL Vp_TPD resulted in 91% mortality after 45 h; co-treatment with 0.1% and 0.2% Aq reduced mortality to ≈12% and ≈6%, respectively, while 1% Aq achieved ≈98% survival. The clinical protection test confirmed that 0.1% Aq preserved high survival across four pathogen inocula (10¹–10⁴ CFU/mL). Conclusively, Aq destabilized the pathogen and therefore transcriptionally silenced multiple virulence determinants, translating into significant in-pond protection for controlling Vp_TPD for shrimp aquaculture.

Obesity is a disorder with an increasing prevalence, associated with cardiovascular and metabolic pathologies. The amount and the distribution pattern of adipose tissue must be considered when managing this disease. Abdominal obesity carries a higher risk of developing obesity-related comorbidities. Body composition methods allow an objective diagnosis and follow-up in obese patients. Although bioelectrical impedance (BIA) is a practical, affordable, and widespread technique to assess body fat percentage (%BF), its accuracy has often been questioned. Therefore, the present study aimed to determine BIA’s accuracy in subjects with abdominal obesity compared to air-displacement plethysmography (AP) as the reference method. Measurements with both body composition techniques were performed on 80 subjects (41 men and 39 women) with a large range of body mass indexes (BMIs). Abdominal obesity, diagnosed as a waist-to-height ratio (WHtR) above 0.5, was present in 28 of them. Agreement between methods was analyzed using t-tests, correlation, and Bland–Altman plots. Compared to AP, BIA underestimated %BF by 3.07 ± 5.81% (CCC = 0.82) in the entire study group. The agreement was comparable in subjects with and without central obesity (2.97 ± 6.21% and 3.26 ± 5.1%, respectively). The changes in body shape geometry due to different adipose tissue distribution patterns only marginally affected BIA’s determinations.

The absence of efficient on-farm interventions against white spot syndrome viral (WSSV) infections can cause significant economic losses to shrimp farmers. With this exploratory study we aimed to test, both in vitro and in vivo, the efficacy of an organic acid mixture (Aq) against WSSV infections in shrimp. In vitro, using shrimp gut primary cells (SGP), 2% Aq significantly reduced WSSV infection and the amounts of H₂O₂ released but had no impact on CAT and SOD expression. In vivo, in a shrimp challenge test, 2% Aq significantly downregulated the expression of proteins involved in WSSV virulence, such as the lipopolysaccharide-β-1,3-glucan-binding protein (LGBP) and the TLR signalling pathway (LvECSIT), and increased the expression of HO-1 oxygenase. Additionally, at 2% Aq, the expression of the digestive-related enzyme carboxypeptidase B was upregulated in the gut, alongside a significant decrease in IL-22 expression, a cytokine usually increased during WSSV infection in shrimp. A low mortality rate (7.33%) was recorded in infected shrimp treated with 2% Aq compared to the 96.66% mortality in the absence of Aq. The peritrophic membrane (PM) was proven essential to ensure Aq efficacy, as the infected and treated PM deficient shrimp (PM−) had a mortality rate of 27.8%, compared to only 9.34% mortality in the infected shrimp at 2% Aq and in the presence of PM (PM+). Aq significantly increased the expression of mucin-1, mucin-2, mucin-5AC, mucin-5B, and mucin-19 in both PM+ and PM− shrimp. Conclusively, organic acid in mixtures can protect farmed shrimp against WSSV infection and increase their survivability through a mediated gut health effect which includes resistance to oxidative stress and improved immunity.

Bioelectrical impedance analysis (BIA) is a widely used and an easy-to-apply method for determining body composition. However, its accuracy depends on population-specific equations. The aim of the present study is to identify the most appropriate fat-free mass (FFM) prediction equation for an Eastern European population using air-displacement plethysmography (ADP) as the reference method. The study group included 101 Caucasian subjects (56 women and 45 men) with an average body mass index (BMI) of 25.37 ± 5.45 kg/m². One set of FFM_BIA values was automatically calculated with the analyzer (Maltron BioScan 920-2), and four others were computed using the published equations of Kyle, Kanellakis, Heitman, and Deurenberg. The results were compared to FFM_ADP values measured using a BOD POD Gold Standard Body Composition Tracking System by Bland–Altman analysis. The smallest bias was obtained with the equation by Deurenberg, which underestimated FFM by only −0.22 ± 4.52 kg. The largest bias was obtained with the equation by Kyle (5.92 ± 4.73 kg), followed by the formula of Kanellakis (3.04 ± 4.65 kg). The equations by Heitman and the Maltron inbuilt formula overestimated FFM by 2.15 ± 4.27 kg and, respectively, 1.95 ± 4.3 kg. Although Maltron’s automatically generated values were very strongly correlated with ADP results (CCC = 0.93, SEE = 4.7), the formula by Deurenberg provided the most reliable estimates in the studied population.