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Lactobacillus delbrueckii, the type species of the genus Lactobacillus, is widely recognized as the primary starter culture in the dairy industry due to its proteolytic activity, which enables it to growth in milk. In this study, a comprehensive genomic analysis of the proteolytic system was conducted on L. delbrueckii strains. The analysis included 27 genomes of L. delbrueckii, with a specific focus on the key enzyme involved in this system, the cell envelope-associated proteinase (CEP). The amino acid sequences, as well as the protein-structure prediction of the CEPs, were compared. Additionally, syntenic analysis of the genomic locus related to the CEPs revealed high conservation in L. delbrueckii subsp. bulgaricus strains, while L. delbrueckii subsp. lactis strains exhibited greater variability, including the presence of insertion sequences, deletions, and rearrangements. Finally, the CEP promoter region and putative regulatory elements responsible for controlling the expression of the proteolytic system in lactobacilli were investigated. Our genomic analysis and in silico characterization of the CEPs contribute to our understanding of proteolytic activity and the potential applications of these lactic acid bacteria in the dairy industry. Further research in this area will expand our knowledge and potential practical uses of these findings.

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Intestinal mucositis promoted by the use of anticancer drugs is characterized by ulcerative inflammation of the intestinal mucosa, a debilitating side effect in cancer patients undergoing treatment. Probiotics are a potential therapeutic option to alleviate intestinal mucositis due to their effects on epithelial barrier integrity and anti-inflammatory modulation. This study investigated the health-promoting impact of Lactobacillus delbrueckii CIDCA 133 in modulating inflammatory and epithelial barrier markers to protect the intestinal mucosa from 5-fluorouracil-induced epithelial damage. L. delbrueckii CIDCA 133 consumption ameliorated small intestine shortening, inflammatory cell infiltration, intestinal permeability, villus atrophy, and goblet cell count, improving the intestinal mucosa architecture and its function in treated mice. Upregulation of Muc2, Cldn1, Hp, F11r, and Il10, and downregulation of markers involved in NF-κB signaling pathway activation (Tlr2, Tlr4, Nfkb1, Il6, and Il1b) were observed at the mRNA level. This work suggests a beneficial role of L. delbrueckii strain CIDCA 133 on intestinal damage induced by 5-FU chemotherapy through modulation of inflammatory pathways and improvement of epithelial barrier function.

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Lactobacillus delbrueckii subsp. lactis CIDCA is a new potential probiotic strain whose molecular basis attributed to the host's benefit has been reported. This study investigated the safety aspects of Lactobacillus delbrueckii subsp. lactis CIDCA 133 based on whole-genome sequence and phenotypic analysis to avoid future questions about the harmful effects of this strain consumption. Genomic analysis showed that L. delbrueckii subsp. lactis CIDCA 133 harbors virulence, harmful metabolites, and antimicrobial resistance-associated genes. However, none of these genetic elements is flanked or located within prophage regions and plasmid sequence. At a phenotypic level, it was observed L. delbrueckii subsp. lactis CIDCA 133 antimicrobial resistance to aminoglycosides streptomycin and gentamicin antibiotics, but no hemolytic and mucin degradation activity was exhibited by strain. Furthermore, no adverse effects were observed regarding mice clinical and histopathological analysis after the strain consumption (5 × 107 CFU/mL). Overall, these findings reveal the safety of Lactobacillus delbrueckii subsp. lactis CIDCA 133 for consumption and future probiotic applications.

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Lactobacillus delbrueckii subsp. lactis CRL 581 beneficially modulates the intestinal antiviral innate immune response triggered by the Toll-like receptor 3 (TLR3) agonist poly(I:C) in vivo. This study aimed to characterize further the immunomodulatory properties of the technologically relevant starter culture L. delbrueckii subsp. lactis CRL 581 by evaluating its interaction with intestinal epithelial cells and macrophages in the context of innate immune responses triggered by TLR3. Our results showed that the CRL 581 strain was able to adhere to porcine intestinal epithelial (PIE) cells and mucins. The CRL 581 strain also augmented the expression of antiviral factors (IFN-α, IFN-ß, Mx1, OAS1, and OAS2) and reduced inflammatory cytokines in PIE cells triggered by TLR3 stimulation. In addition, the influence of L. delbrueckii subsp. lactis CRL 581 on the response of murine RAW macrophages to the activation of TLR3 was evaluated. The CRL 581 strain was capable of enhancing the expression of IFN-α, IFN-ß, IFN-γ, Mx1, OAS1, TNF-α, and IL-1ß. Of note, the CRL 581 strain also augmented the expression of IL-10 in macrophages. The results of this study show that the high proteolytic strain L. delbrueckii spp. lactis CRL 581 was able to beneficially modulate the intestinal innate antiviral immune response by regulating the response of both epithelial cells and macrophages relative to TLR3 activation.

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Lactobacillus delbrueckii subsp. lactis CIDCA 133 (CIDCA 133) has been reported as a potential probiotic strain, presenting immunomodulatory properties. This study investigated the possible genes and molecular mechanism involved with a probiotic profile of CIDCA 133 through a genomic approach associated with in vitro and in vivo analysis. Genomic analysis corroborates the species identification carried out by the classical microbiological method. Phenotypic assays demonstrated that the CIDCA 133 strain could survive acidic, osmotic, and thermic stresses. In addition, this strain shows antibacterial activity against Salmonella Typhimurium and presents immunostimulatory properties capable of upregulating anti-inflammatory cytokines Il10 and Tgfb1 gene expression through inhibition of Nfkb1 gene expression. These reported effects can be associated with secreted, membrane/exposed to the surface and cytoplasmic proteins, and bacteriocins-encoding genes predicted in silico. Furthermore, our results showed the genes and the possible mechanisms used by CIDCA 133 to produce their beneficial host effects and highlight its use as a probiotic microorganism.

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Raw meat emulsions may have natural, spoilage and pathogenic microorganisms due to the origin and characteristics of this food matrix. All of these microorganisms must be minimized during industrial processing to make food consumption safe and meet quality regulations. Therefore, in this research, the effect of probe ultrasound on the inactivation of three kinds of microorganisms in a raw meat emulsion is evaluated. The microorganisms are: natural microflora NAM, Listeria monocytogenes LIS, and Lactobacillus delbrueckii LAC. A high-intensity probe ultrasound system was used, during 1.0, 2.5, 5.0, 7.5 and 10 min, with pulsed waves of 0.0, 10, 20 and 30 seg, and 200, 250, 300, 350 and 400 W of power. The interrelation between time, wave pulse cycle, and power factors was assessed. The results showed a positive linear independence effect in the treatments without wave pulse for each microorganism, and a quadratic interaction with the time and the ultrasound power for the inactivation of the three kinds of microorganisms. Besides, the desirability function for the inactivation reached up to 60% of the microbial population with the probe ultrasound treatment, with 10 min, a 7.56 s wave pulse and 400 W of power. Thus, these results could be useful to decide the incorporation of mild and emerging technologies in a meat industry line process.

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ABSTRACT: The study aimed to analyze the physicochemical properties and starter culture viability of frozen yogurts produced with liquid cheese whey (LCW) and inulin at different proportions (F0: 66% LCW and 0% inulin; F1: 65% LCW and 1% inulin; F2: 64% LCW and 2% inulin; F3: 62% LCW and 4% inulin). Results demonstrated that the frozen yogurt F3 presented higher total solids and carbohydrates levels. LCW and inulin contributed to the overrun increase (11.8-18.2%) but did not interfere significantly in the retardation of the melting rate and range in the samples' hardness. Over the storage time, formulation F3 showed lower pH and higher titratable acidity. However, from the sixtieth day of storage, the formulations of frozen yogurts varied in the pH and titratable acidity profile associated with the decline in the viability of starter cultures. Even so, the inulin supplementation positively affected the strains' viability during storage. Based on our data, the formulation F3 presented better nutritional value, physicochemical characteristics, and stability over the storage period.

RESUMO: O estudo teve como objetivo analisar as propriedades físico-químicas e a viabilidade das culturas starters de frozen yogurts produzidos com soro líquido de queijo (SLQ) e inulina em diferentes proporções (F0: 66% SLQ e 0% inulina; F1: 65% SLQ e 1 % inulina; F2: 64% SLQ e 2% inulina; F3: 62% SLQ e 4% inulina). Os resultados demonstraram que o frozen yogurt F3 apresentou maiores teores de sólidos totais e carboidratos. SLQ e inulina contribuíram para o aumento do overrun (11.8-18.2%), mas não interferiram significativamente no retardo da taxa de derretimento e na variação da dureza das amostras. Ao longo do tempo de armazenamento, a formulação F3 apresentou menor pH e maior acidez titulável. Porém, a partir do sexagésimo dia de armazenamento, as formulações de frozen yogurts variaram o perfil de pH e acidez titulável associado ao declínio da viabilidade das culturas starter. Mesmo assim, a suplementação de inulina afetou positivamente a viabilidade das cepas durante o armazenamento. Com base em nossos dados, a formulação F3 apresentou melhor valor nutricional, características físico-químicas e estabilidade ao longo do período de armazenamento.

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The study aimed to analyze the physicochemical properties and starter culture viability of frozen yogurts produced with liquid cheese whey (LCW) and inulin at different proportions (F0: 66% LCW and 0% inulin; F1: 65% LCW and 1% inulin; F2: 64% LCW and 2% inulin; F3: 62% LCW and 4% inulin). Results demonstrated that the frozen yogurt F3 presented higher total solids and carbohydrates levels. LCW and inulin contributed to the overrun increase (11.8-18.2%) but did not interfere significantly in the retardation of the melting rate and range in the samples’ hardness. Over the storage time, formulation F3 showed lower pH and higher titratable acidity. However, from the sixtieth day of storage, the formulations of frozen yogurts varied in the pH and titratable acidity profile associated with the decline in the viability of starter cultures. Even so, the inulin supplementation positively affected the strains’ viability during storage. Based on our data, the formulation F3 presented better nutritional value, physicochemical characteristics, and stability over the storage period.

O estudo teve como objetivo analisar as propriedades físico-químicas e a viabilidade das culturas starters de frozen yogurts produzidos com soro líquido de queijo (SLQ) e inulina em diferentes proporções (F0: 66% SLQ e 0% inulina; F1: 65% SLQ e 1 % inulina; F2: 64% SLQ e 2% inulina; F3: 62% SLQ e 4% inulina). Os resultados demonstraram que o frozen yogurt F3 apresentou maiores teores de sólidos totais e carboidratos. SLQ e inulina contribuíram para o aumento do overrun (11.8-18.2%), mas não interferiram significativamente no retardo da taxa de derretimento e na variação da dureza das amostras. Ao longo do tempo de armazenamento, a formulação F3 apresentou menor pH e maior acidez titulável. Porém, a partir do sexagésimo dia de armazenamento, as formulações de frozen yogurts variaram o perfil de pH e acidez titulável associado ao declínio da viabilidade das culturas starter. Mesmo assim, a suplementação de inulina afetou positivamente a viabilidade das cepas durante o armazenamento. Com base em nossos dados, a formulação F3 apresentou melhor valor nutricional, características físico-químicas e estabilidade ao longo do período de armazenamento.

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The study aimed to analyze the physicochemical properties and starter culture viability of frozen yogurts produced with liquid cheese whey (LCW) and inulin at different proportions (F0: 66% LCW and 0% inulin; F1: 65% LCW and 1% inulin; F2: 64% LCW and 2% inulin; F3: 62% LCW and 4% inulin). Results demonstrated that the frozen yogurt F3 presented higher total solids and carbohydrates levels. LCW and inulin contributed to the overrun increase (11.8-18.2%) but did not interfere significantly in the retardation of the melting rate and range in the samples hardness. Over the storage time, formulation F3 showed lower pH and higher titratable acidity. However, from the sixtieth day of storage, the formulations of frozen yogurts varied in the pH and titratable acidity profile associated with the decline in the viability of starter cultures. Even so, the inulin supplementation positively affected the strains viability during storage. Based on our data, the formulation F3 presented better nutritional value, physicochemical characteristics, and stability over the storage period.(AU)

O estudo teve como objetivo analisar as propriedades físico-químicas e a viabilidade das culturas starters de frozen yogurts produzidos com soro líquido de queijo (SLQ) e inulina em diferentes proporções (F0: 66% SLQ e 0% inulina; F1: 65% SLQ e 1 % inulina; F2: 64% SLQ e 2% inulina; F3: 62% SLQ e 4% inulina). Os resultados demonstraram que o frozen yogurt F3 apresentou maiores teores de sólidos totais e carboidratos. SLQ e inulina contribuíram para o aumento do overrun (11.8-18.2%), mas não interferiram significativamente no retardo da taxa de derretimento e na variação da dureza das amostras. Ao longo do tempo de armazenamento, a formulação F3 apresentou menor pH e maior acidez titulável. Porém, a partir do sexagésimo dia de armazenamento, as formulações de frozen yogurts variaram o perfil de pH e acidez titulável associado ao declínio da viabilidade das culturas starter. Mesmo assim, a suplementação de inulina afetou positivamente a viabilidade das cepas durante o armazenamento. Com base em nossos dados, a formulação F3 apresentou melhor valor nutricional, características físico-químicas e estabilidade ao longo do período de armazenamento.(AU)

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Proteolytic starter cultures with intrinsic immunomodulatory activities are desirably features for the development of functional foods, which would significantly reduce the cost of their production (one-strain starter) having an additional beneficial effect on the host. In this work, Lactobacillus delbrueckii strains were selected according to their ability to efficiently hydrolyse ß-casein and to modulate the immune system. Among 36 strains evaluated, the highest proteolytic activities were found for L. delbrueckii subsp. lactis CRL581 and L. delbrueckii subsp. bulgaricus CRL656. The immunomodulatory effect of both strains and their ß-casein hydrolysates (CRL581 and CRL656 hydrolysates, respectively) were studied in a murine model. Balb/c mice were fed lactobacilli or their hydrolysates for three days. One day after the last lactobacilli or hydrolysate treatments, mice were challenged with the Toll-like receptor 3 (TLR3) agonist poly(I:C) by intraperitoneal injection. Before and after poly(I:C) challenge the phagocytic and microbicidal activity of peritoneal macrophages, intestinal immunoglobulin A (IgA), cytokine profile, and histological analysis of the intestine were analysed. L. delbrueckii subsp. lactis CRL581 significantly increased the activation of peritoneal macrophages as well as the levels of intestinal IgA, interleukin (IL)-10 and interferon (IFN)-γ when compared to untreated controls. In addition, the CRL581 strain was able to significantly reduce the intestinal inflammatory damage triggered by TLR3 activation. L. delbrueckii CRL581 increased the levels of IL-10, IFN-γ and IFN-ß, and reduced tumour necrosis factor alpha and IL-6 concentrations in the intestine of poly(I:C)-challenged mice. No immunomodulatory effects were observed for the CRL656 strain or for the CRL581 or CRL656 hydrolysates. The results of this work show that the technologically relevant and high proteolytic strain L. delbrueckii CRL581 is able to beneficially modulate the intestinal innate antiviral immune response. Although further studies with the CRL581 strain are required to corroborate and deepen its immunological effects, this bacterium is an interesting alternative for the development of new functional foods with antiviral capabilities.

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Hypertension is one of the main risk factors for cardiovascular disease. Functional foods containing bioactive peptides have been proposed as a strategy to decrease blood pressure (BP) in subjects under no pharmacological treatment. The aim of this study was to compare the effect of low-sodium, low-fat (LSLF) cheese and LSLF cheese containing Lactobacillus delbrueckii subsp. lactis CRL 581 (LSLF581) on BP in pre-hypertensive and stage 1 hypertensive subjects. Sixty-one pre-hypertensive and stage 1 hypertensive subjects assigned to one of twos (LSLF, n= 29 and LSLF581, n= 32) participated in this 12-month prospective, randomized, double-blind, crossover trial. Twenty-four h ambulatory BP monitoring was performed at the beginning and at the end of each four-week study period. Systolic and diastolic BP decreased in both study groups, but differences between groups were not significant (systolic, -1.78 and -0.2 mmHg; diastolic, -1.54 and -0.42 mmHg in LSLF581 and LSLF, respectively). Although our results could not support a BP lowering effect of LSLF581, small BP reductions could favorably prevent cardiovascular disease development.

La hipertensión arterial es uno de los principales factores de riesgo de enfermedad cardiovascular. Los alimentos funcionales que contienen biopéptidos constituyen una estrategia útil para disminuir la presión arterial (PA) en personas que no están bajo tratamiento farmacológico. El objetivo del estudio fue comparar el efecto de un queso bajo en sodio y bajo en grasas (BSBG) y el mismo queso con Lactobacillus delbrueckii subsp. lactis CRL 581 (BSBG581) sobre la PA en personas con prehipertensión y estadio 1 de hipertensión arterial. Realizamos un estudio prospectivo, randomizado, cruzado y doble ciego durante 12 meses en 61 personas con prehipertensión y estadio 1 de hipertensión arterial, asignadas a dos grupos: BSBG (n= 29) y BSBG581 (n= 32). Se realizó monitoreo ambulatorio de la PA (MAPA) durante 24 h al comienzo y al final de cada etapa del estudio (cuatro semanas). La PA sistólica y diastólica disminuyó en ambos grupos, aunque las diferencias entre grupos no fueron significativas (sistólica, -1.78 y -0.2 mmHg; diastólica -1.54 y -0.42 mmHg en BSBG581 y BSBG respectivamente). Aunque nuestros resultados no pueden confirmar el efecto hipotensor del queso BSBG581, las reducciones moderadas de la PA podrían prevenir el desarrollo de enfermedad cardiovascular.

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El objetivo principal de esta investigación fue determinar las condiciones adecuadas de crecimiento del Lactobacillus delbrueckii subsp. bulgaricus y Streptococcus thermophilus para la producción de ácido láctico, utilizando como sustrato lactosuero proveniente del departamento del Cesar, Colombia. El ácido láctico es producto de la extracción y purificación de caldos de fermentación donde se usa como bacterias fermentativas el co-cultivo Lactobacillus delbrueckii subsp. bulgaricus y Streptococcus thermophilus, usualmente usadas para la producción de yogurt. El sustrato es suplementado con extracto de levadura, fosfato de amonio como fuente de nitrógeno y carbonato de calcio como neutralizante. Esto con el fin de optimizar el consumo, por parte de las bacterias, del carbohidrato principal presente en el suero (lactosa). Durante la fermentación se controló la concentración del inóculo y temperatura, el tiempo máximo fue 72 h. La purificación se llevó a cabo mediante esterificación, filtración de sólidos formados con la reacción y extracción de agua por evaporación y posterior arrastre con nitrógeno. Finalmente se obtuvo ácido láctico con un 78,0% de pureza (36,7 g/L), el cual se caracterizó a través de espectroscopía infrarroja.

O principal objetivo desta pesquisa foi determinar as condições adequadas de crescimento do Lactobacillus delbrueckii subsp. bulgaricus e Streptococcus thermophilus para a produção de ácido láctico utilizando como substrato soro de leite obtido do Departamento do Cesar, Colômbia. O ácido láctico é um produto da extração e purificação de caldos de fermentação, no qual são usadas como bactérias fermentativas Lactobacillus delbrueckiisubsp. bulgaricus e Streptococcus thermophilus, normalmente usadas para a produção de iogurte. O substrato é suplementado com extrato de levedura, fosfato de amonio como fonte de nitrogênio e de carbonato de cálcio como um neutralizador, para optimizar o consumo, pelas bactérias, do principal hidrato de carbono presente no soro (lactose). Durante a fermentação os seguintes fatores foram controlados: concentração do inóculo e temperatura, e atingiu num máximo de 72 h. A purificação foi efetuada por esterificação, filtração dos sólidos formados com a reação e remoção de água por evaporação e varrimento com nitrogênio. Finalmente o ácido láctico foi obtido com 78,0% de pureza (36,7 g/L), e foi caracterizado por espectroscopia de infravermelho.

The main objective of this research was to determine the proper growth conditions of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus for the production of lactic acid using serum as substract. This serum was obtain from the department of Cesar, Colombia. Lactic acid is the result of the extraction and purification of fermentation broths in which bacteria Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus are used, which are usually used for the production of yogurt. The substrate was supplemented with yeast extract, ammonium phosphate as a nitrogen source, and calcium carbonate as a neutralizer, in order to optimize the consumption, by the bacteria, of the main carbohydrate present in serum (lactose). During the fermentation (up to 72 h) the inoculums concentration, and temperature were controlled. Purification consisted in esterification, filtration of solids formed during the reaction, and removing of water by evaporation and nitrogen influx. Finally, lactic acid was obtained with 78,0% purity (36.7 g/L), which was characterized by infrared spectroscopy.

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Poly-ß-hydroxybutyrate (PHB) is a biodegradable intracellular microbial product produced by many bacteria and it is comparable to some of the petrochemical derived thermoplastics such as polypropylene. One of the main barriers for the commercial exploitation is the high cost of the substrate for the production of biopolymer. The utilization of mixed microbial cultures facilitates the use of complex substrates thereby reducing the cost of PHB production. In the present study, mixed culture systems were evaluated for PHB production. Bacillus firmus NII 0830 was used for the production of PHB since it accumulates a large amount of PHB and a second organism Lactobacillus delbrueckii NII 0925 was used to provide lactic acid. FTIR and 1H NMR analyses revealed that the PHB extracted from pure culture and mixed culture showed exact match to that of standard PHB. Biodegradation studies of the PHB blends showed 87% degradation. It was also found that a consortium of organisms degraded the films faster than a single organism.

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Lactobacillus delbrueckii subsp. bulgaricus CIDCA 333 was dehydrated on desiccators containing silica gel in the presence of 20% w/w of two types of galacto-oligosaccharides (GOS Biotempo and GOS Cup Oligo H-70®) and lactulose, until no changes in water desorption were detected. After rehydration, bacterial growth was monitored at 37°C by determining: (a) the absorbance at 600 nm and (b) the near infrared spectra (NIR). Principal component analysis (PCA) was then performed on the NIR spectra of samples dehydrated in all conditions. A multiparametric flow cytometry assay was carried out using carboxyfluorescein diacetate and propidium iodide probes to determine the relative composition of damaged, viable, and dead bacteria throughout the growth kinetics. The absorbance at 600 nm and the position of the second derivative band at ∼1370 nm were plotted against the time of incubation. The efficiency of the protectants was GOS Biotempo > GOS Cup Oligo H-70® > lactulose. The better protectant capacity of GOS Biotempo was explained on the basis of the lower contribution of damaged cells immediately after rehydration (t = 0). PCA showed three groups along PC1, corresponding to the lag, exponential and stationary phases of growth, which explained 99% of the total variance. Along PC2, two groups were observed, corresponding to damaged or viable cells. The results obtained support the use of NIR to monitor the recovery of desiccated microorganisms in real time and without the need of chemical reagents. The use of GOS and lactulose as protectants in dehydration/rehydration processes was also supported.

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This study aimed to assess the transit tolerance of potential probiotic dairy Lactobacillus strains in human uppergastrointestinal tract in vitro, and to evaluate the effect of EPS production on the viability and adhesion of these strains. Survival and adhesion of two exopolysaccharide (EPS)-producing L. delbrueckii subsp. bulgaricus strains (B3 and B2) and E. coli ATCC11229 were assessed after the exposure of different pH (gastric juice) and gastric plus pancreatic juice challenges. In the artificial gastric juice (pH 2), both the viability of the strain B3 and B2 was decreased. Artificial juice treatments significantly reduced the adhesion to caco-2 cells (P< 0.05). High EPS-producing B3 survived better in the adverse gastrointestinal conditions and showed better ability of adhesion to Caco-2 cells when assessed for competition with E. coli ATCC 11229 compared to low EPS-producing B2. This investigation showed that EPS production could be affected or be involved in the viability, adherence and competition of L. delbrueckii subsp. bulgaricus strains and support the potential of B3 strain for development of new probiotic products.

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Exopolysaccharides (EPSs) production was studied by Lactobacillus delbrueckii subsp. bulgaricus (B3, G12) and Streptococcus thermophilus (W22) in the medium containing various carbon sources (glucose, fructose, sucrose or lactose). For all the strains, glucose was the most efficient carbon source and B3, G12 and W22 strains produced 211, 175 and 120 EPS mg/L respectively. Also, the influence of different concentrations of glucose (5,10,15,20,25,30 g/L) on EPS production and growth was studied. The results indicated that EPS production and growth were stimulated by the high glucose concentration (30 g/L).