Survival of commercial probiotic strains and their effect on dark chocolate synbiotic snack with raspberry content during the storage and after simulated digestion

BACKGROUND: A key challenge for manufacturers of pro-health food containing active probiotic microorganisms is to develop a product with attractive sensory features along with maintenance of declared number of microorganisms during storage and transfer by alimentary tract. RESULTS: The highest concentration of polyphenols was observed in snacks without an additive of probiotics as well as those with an additive of L. rhamnosus and B. animalis bacteria and concentration of these compounds increased by 9.5% during six months of storage. None of the products distinguished itself in the sensorial assessment although each was assessed positively. The number of microorganisms was stable and comparatively high during six months of storage at a room temperature and in cooling conditions (108 cfu/g). In the digestion model, an influence of aggressive digestion conditions was examined in the alimentary tract on the number of microorganisms, which allowed to arrange strains from the most resistant (S. boulardii) to the most sensitive (B. breve). It must be noted that currently on the market there is no available snack containing probiotic yeast as well as there is no literature data on works on such formulation of food. CONCLUSIONS: In the newly developed snack made of chocolate, in which sugar has been replaced with maltitol, a raw material was added in the form of raspberry, prebiotic in the form of inulin and a strain of probiotic bacteria, including the unprecedented so far S. boulardii, which stands a high chance to occupy a good place on the market of functional food.

Inhibition of Carbapenem-Resistant NDM-1 Klebsiella pneumoniae isolated from a Hospital Outbreak Patient by a Synbiotic: A Nonantibiotic Treatment Option

Objective: Klebsiella (K.) pneumoniae is globally responsible for an alarming increase in hospital infections, especially in intensive care units (ICUs). The acquisition of resistance against a broad range of antibiotics has turned infections with this pathogen into a major worldwide healthcare concern. The aim of the study was to investigate if multistrain synbiotics can complement the current treatment options of multidrug-resistant K. pneumoniae infections. Methods: Antimicrobial susceptibility and PCR testing were used to characterize the K. pneumoniae causing a hospital outbreak. Effect of multistrain synbiotic administration on the presence of K. pneumoniae in an infected patient was investigated by microbiological testing for the pathogen. Effects of the synbiotic mixture and its individual probiotic bacteria on K. pneumoniae isolated from patients and of the K. pneumoniae subsp. pneumoniae ATCC© 700603TM reference strain were investigated by pathogen in-vitro inhibition experiments. Results: The outbreak K. pneumoniae strain was found to be resistant against a range of antibiotics including carbapenems, and to be a producer of New Delhi metallo-β-lactamase 1 (NDM-1). Treatment of a NDM-1 K. pneumoniae carrier with a multistrain synbiotic resulted in successful elimination of the pathogen from the patient. In-vitro inhibition experiments showed that the NDM-1 K. pneumoniae (and the reference strain K. pneumoniae subsp. pneumoniae, ATCC© 700603TM) could be effectively inhibited by the bacteria mixture of the synbiotic preparation. Conclusion: Findings of the study indicate for the first time that a multistrain synbiotic can add to the treatment repertoire available for the management of NDM-1 K. pneumoniae infections.

Aflatoxin M1 Reduction in Milk by a Novel Combination of Probiotic Bacterial and Yeast Strains

Milk and milk products are an important contribution to the human diet especially children. However, the presence of aflatoxins as AFM1 in milk and milk products are considered undesirables due to their health risks in consumer's body. For that reason, this study aimed to assess the ability of some microbial species on aflatoxin removal especially the AFM1 in the milk. The AFM1 residue was determined by HPLC after different incubation period (12 h, 24 h, 48 h and 72 h) of the probiotic bacteria and yeasts in PBS as a model for AFM1 determination. This study was performed during the period between 2015 and 2017. The combination of nonviable probiotic bacterial and yeast species (Lactobacillus Plantarum, Lactobacillus acidophilus, Bifidobacterium bifidum, Kluyveromyces lactis and Saccharomyces cerevisiae) succeeded to reduce AFM1 from 50 (ng/ml) during the incubation periods; 12 h, 24 h, 48 h and 72 h, into 9.72±1.31, 6.68±0.55, 5.70±0.33 and 4.56±0.15 ng ml-1, respectively. The highest AFM1 removal % was recorded as; 80.56%, 86.64%, 88.60% and 90.88% in the treated milk samples in a respective manner. Sensor evaluation was carried out using Yoghurt as a model in sample size 50 gm for each sample. Conclusion: This study concluded that the combination consisting of probiotic bacteria and yeasts could be used in reducing the concentration of the AFM1 in aflatoxin contaminated milk.

Antithrombotic Activity of Milk Protein Hydrolysates by Lactic Acid Bacteria Isolated from Commercial Fermented Milks

ABSTRACT In the last decade, thrombosis has been one of the pathologies with high incidence creating great concern in Health Institutes all around the world. Considering this, the aim of this research was to determine the antithrombotic activity of peptides released during lactic fermentation. Reconstituted skim milk powder was fermented by Lactobacillus casei Shirota and Lactobacillus johnsonii LA1 isolated from commercial fermented milks. The hydrolysis degree and proteolytic profile were analyzed by trinitrobenzenesulfonic acid spectrophotometry method and by peptide polyacrylamide electrophoresis gel separation. The milk fermented by Lactobacillus casei Shirota exhibited a higher concentration of free amino groups than that fermented by Lactobacillus johnsonii LA1. Additionally, in both fermentation systems peptides with molecular weights lower than 1.4 kDa were observed. The highest inhibition of thrombin (31.67±2.35%) was observed in milk fermented by Lactobacillus johnsonii LA1 at 10 hours of fermentation. Finally, no relationship was found between the antithrombotic capacity and the proteolytic profile.

Isolation of Lactic Acid Bacteria from Ewe Milk, Traditional Yoghurt and Sour Buttermilk in Iran.

A total of 63 samples including ewe milk, yoghurt and traditional buttermilk were collected from Myaneh and Hashrood (Azarbayjan-e-Sharqi, Iran) and screened for the presence of Lactic Acid Bacteria (LAB). Based on routine cultural characteristic, general morphological and biochemical assay, 77 out of 168 bacterial isolates were identified as LAB. These isolates were examined for the presence of inhibitory activity against other randomly selected LAB isolates. Thirty-three strains showed antagonistic activity against the closely related LAB strains and were further challenged against other gram-positive and gram-negative pathogens including Listeria monocytogenes, Staphylococcus aureus and Salmonella entritidis. Based on their zones of inhibition diameters the isolates showing maximum inhibitory activity against these pathogens were selected for detailed investigations. The selected isolates were identified to species level by 50CHL API system and were challenged to heat, acid and bile salt. Most of the strains were able to survive at different pH ranges, while one strain of Pedicoccus acidilactici and Lactobacillus paracasei were able to tolerate all ranges of pH during 24 h of incubation. In addition, Lactobacillus brevis was found as the most resistant strain being able to resist all concentrates of bile after 4 h. The results indicated the probiotic potential of the isolates, as majority of the selected LAB isolates were capable of resisting high temperatures, acidic pH values and bile concentrations of 0.7%.

Potencial de biodegradação de microcistinas por microrganismos / Biodegradation potential of microcystins by microorganisms

Para minimizar os problemas relacionados à ocorrência de cianobactéria em águas destinadas ao consumo humano há necessidade de se realizar estudos de alternativas técnicas de tratamento com destaque aos biofilmes com potencial de degradação de microcistinas (MC). O presente trabalho teve como objetivo avaliar o potencial de degradação de MC pela bactéria Sphingosinicella microcystinivorans B9, diferentes cepas de leveduras e bactérias probióticas. O teste foi efetuado com extrato de MC e diferentes quantidades de biovolume e densidade celular dos microrganismos. Os tratamentos foram mantidos a 27ºC com rotação de 100 rpm e as amostras para análise de MC e contagem dos microrganismos foram retiradas após 0 e 96 horas de contato. A bactéria B9 apresentou maior degradação de MC, chegando a 98% após 96 horas.

To minimize problems related to the occurrence of cyanobacteria in water for human consumption there is need to investigate alternative treatment techniques with emphasis on biofilms with the potential degradation of microcystins (MC). This study aimed to evaluate the potential degradation of MC by bacteria Sphingosinicella microcystinivorans B9, different strains of yeast and probiotic bacteria. The test was carried out with the extract obtained from strain Microcystis sp. In the tests biomass and cultures of microorganisms were used and the treatments were maintained at 27ºC with 100 rpm. Samples for analysis of MC and for counting the microorganisms were collected at 0 and 96 hours. The bacterium B9 presented the highest potential of degradation of MC reaching 98% after 96 hours.

Bacteriocin-producing enterococci from rabbit meat

Aims: Enterococci are lactic acid bacteria belonging to the division Firmicutes. They occur in different ecosystems, rabbits including. Enterococci can possess probiotic properties and produce antimicrobial substances-bacteriocins. Rabbit meat as nutritionally healthy food offers novel source to study bacteriocin-producing and/or probiotic enterococci. Methodology and results: Enterococci were detected from rabbit meat samples (42). Most of the isolates were allotted to the species Enterococcus faecium by PCR method. The isolates have possessed the structural genes for enterocins A, P, B production. The inhibitory substances produced by the isolated enterococci inhibited the growth of 12 indicators. Of 34 isolates, 15 strains have shown the antimicrobial activity against L. monocytogenes CCM 4699, 12 strains against S. aureus 3A3, 10 strains against S. aureus 5A2 as well as Salmonella enterica serovar Enteritidis PT4. Moreover, enterococci have tolerated 5 % bile, low pH; they have produced lactid acid in the amount from 0.740 ± 0.091 to 1.720 ± 0.095 mmol/l. The isolates were mostly sensitive to antibiotics. Conclusion, significance and impact of study: Bacteriocin-producing strain E. faecium M3a has been selected for more detail characterization of its bacteriocin and probiotic properties with the aim for its further application as an additive.

Efecto de bacterias probióticas en el cultivo larvario del ostión de placer Crassostrea corteziensis (Bivalvia: Ostreidae)

Effect of probiotic bacteria on survival and growth of Cortez oyster larvae, Crassostrea corteziensis (Bivalvia: Ostreidae). Disease control problems have major constraints in aquaculture production, and the use of probiotics in larviculture is a valid alternative to antibiotics. This study analyzed the effect of probiotic bacteria on survival and final size of Cortez oyster larvae Crassostrea corteziensis. Two different probiotic concentrations were evaluated, 1x10(4) and 1x10(5)CFU/ml of Lactic acid bacteria (strain NS61) isolated from Nodipecten subnodosus, and bacilli isolated from the white leg shrimp, Litopenaeus vannamei (Pseudomonas aeruginosa, strain YC58) and C. corteziensis (Burkholderia cepacia, strain Y021). Bacteria were added directly into culture tanks, starting the bioassays from veliger to pediveliger stages as follows: (1) Control, without probiotics; (2) lactic acid bacteria (Lb); (3) bacilli mix (Mb) in a proportion 1:1. Results showed a higher larval survival with Lb and Mb at a dose of 1x10(4)CFU/ml compared to the control group. Larvae exposed to Mb at 1x10(5)CFU/ml showed higher survival than Lb and control. Larval final size was not significantly increased with the tested probiotics, but larvae treated with Lb at 1x105CFU/ml showed less survival rate than those treated at 1x10(4)CFU/ml. This study showed the beneficial effect of these probiotics, added individually or mixed in C. corteziensis larvae culture. Rev. Biol. Trop. 59 (1): 183-191. Epub 2011 March 01.

El ostión de placer u ostra del Cortés (Crassostrea corteziensis) se considera como una especie con potencial para ser cultivada en gran escala. Sin embargo, al igual que en otros bivalvos, la alta mortalidad que se presenta durante la etapa larvaria y juvenil, es el principal problema que limita el desarrollo del cultivo en el laboratorio. Un método que está ganando aceptación en la acuicultura es el uso de bacterias probióticas para controlar patógenos microbianos. Este estudio analiza el efecto de estas bacterias en la supervivencia y talla final de larvas de ostión de placer Crassostrea corteziensis. Se utilizó una cepa de bacterias ácido lácticas (cepa NS61) aisladas N. subnodosus, así como de bacilos aislados de L. vannamei (Pseudomonas aeruginosa, cepa YC58) y de C. corteziensis (Burkholderia cepacia, cepa Y021). Las cepas se evaluaron por inmersión en cultivos larvarios de C. corteziensis a dos concentraciones diferentes, hasta completar el estadio pediveliger. Los organismos se trataron con bacterias ácido lácticas (Lb), una mezcla de bacilos (Lb) en proporción 1:1 y un grupo control. La concentración de 1x10(4)UFC/ml registró una mayor supervivencia con Lb y Mb respecto al grupo control. La supervivencia con Mb a una concentración de 1x10(5)UFC/ml fue mayor que la del grupo control y del grupo tratado con Lb. Los resultados mostraron que las larvas de C. corteziensis tratadas con probióticos no incrementaron significativamente su talla respecto a las larvas del grupo control. Mientras que las tratadas con Lb a la concentración mayor, 1x10(5)UFC/ml, mostraron una disminución de la supervivencia respecto a las tratadas con 1x10(4)UFC/ml. Este estudio demostró el efecto benéfico de cepas probióticas utilizadas individualmente o en mezcla en el cultivo larvario de C. corteziensis.

Identificación, cuantificación y determinación del perfil de sensibilidad a antibióticos de bacterias prebióticas adicionadas a productos de consumo frecuente en Costa Rica / Identification, quantification and antimicrobial susceptibility pattern of probiotic bacteria added to common use food products in Costa Rica

En los últimos años, debido a la alta demanda de los productos adicionados con probióticos y los múltiples beneficios nutricionales y terapéuticos asociados, la investigación sobre estos microorganismos ha progresado considerablemente, se han realizado avances notables en su selección y caracterización. De manera general, diversas entidades como la Organización Mundial de la Salud (OMS) y la Organización de las Naciones Unidas para la agricultura y la alimentación (FAO) recomiendan que se declare en la etiqueta del producto, tanto la especie como la cantidad de microorganismos probióticos viables presentes. En este trabajo se analizaron seis productos adicionados con probióticos, con el fin de evaluar su concentración a lo largo de la vida útil del producto, se identificaron las cepas aisladas para corroborar la información declarada en la etiqueta y se determinó su perfil de susceptibilidad a antibióticos. Como resultado del análisis se encontró que las cepas adicionadas a los productos evaluados se mantienen en concentraciones aceptables durante los 28 días de vida útil de los productos. La identificación de las cepas aisladas no coincidió, en varios casos, con la estipulada por la etiqueta, no obstante, el método utilizado se basa en la capacidad de fermentar carbohidratos y no en pruebas genotípicas. Con respecto a los perfiles de susceptibilidad encontrados para las cepas analizadas, son necesarios estudios adicionales que establezcan la naturaleza intrínseca o adquirida de los determinantes de resistencia, y que evidencien si estos están codificados en elementos transferibles entre bacterias.

In the last years, due to the high demand of food products supplemented with probiotics and the multiple nutritional and therapeutic benefits associated with them, research on these microorganisms has advanced considerably, including their selection and characterization. As a general recommendation, several entities as World Health Organization (WHO) and United Nations Organization for Agriculture and Food recommend that the specification of the alive species contained and their number shall appear in the label of the product. In the present study, six different commercially available products, supplemented with probiotics were analyzed, in order to evaluate the concentration of microorganisms through the shelf life of the product, identify the strains isolated and determine the antibiotic susceptibility pattern of these. Results demonstrated that the strains isolated kept acceptable concentrations during the 28 days of storage. Nevertheless, the identification of these strains variated from the one reported on the label on several of the products tested. This can be due to the commercial method used for the identifications, which is based in the carbohydrate fermentation pattern and not in genotypic trials. The antimicrobials’ susceptibility patterns found show that further research shall be performed in order to establish the intrinsic or acquired nature of the resistance determinants, and if these are codified by transferable elements among bacteria.

Study on the Probiotic Properties of Some Lactic Acid Bacteria Strains / 微生物学通报

The acid-and bile-tolerant,the transit tolerance,the ability to adhere to human intestinal cells and the ability to assimilate cholesterol of seven lactic acid bacteria and Bifidobacteria strains were studied by an in vitro method.The abilities to adhere to CCL-187 cells were different between all strains tested(P95% when incubated in simulated gastric and small intestinal juice for 3h and 4h respectively.All strains have the ability to grow in the presence of bile,and the ability varied among the strains.The cholesterol assimilated ranged from 25% to 54%.Enterococcus faecium A30,A31 and Lactobacillus acidophilus A878,PB1,which assimilated more than 40% cholesterol from circumstances,can be good hypocholesterolemic candidate strains for functional food.