scholarly journals Extracellular and intracellular DNA for bacterial profiling of long-ripened cheeses

2020 ◽  
Vol 367 (13) ◽  
Author(s):  
Tommaso Bardelli ◽  
Lia Rossetti ◽  
Miriam Zago ◽  
Domenico Carminati ◽  
Giorgio Giraffa ◽  
...  
Keyword(s):  
Lactobacillus Rhamnosus ◽  
Streptococcus Thermophilus ◽  
Lactobacillus Helveticus ◽  
Extracellular Dna ◽  
Intact Cells ◽  
Additional Information ◽  
Buffer Solutions ◽  
Novel Approach ◽  
Three Samples ◽  
Bacterial Profiling

ABSTRACT A novel approach was developed to extract the extracellular DNA (eDNA), i.e. the free DNA outside the microbial cell, compared to the intracellular DNA (iDNA). The two DNA fractions were investigated in seven long-ripened cheeses. Among different buffer solutions tested, EDTA 0.5 M at pH 8 enabled a mild homogenization of cheese samples and the highest eDNA recovery. The extraction protocol was tested on single strains of lactic acid bacteria characterizing many Italian long-ripened cheeses, such as Streptococcus thermophilus, Lactobacillus helveticus, and Lactobacillus rhamnosus. The method resulted suitable for eDNA extraction because it minimized cell-lysis, avoiding the leakage of iDNA from the cells. The yields of eDNA, ranging from 0.01 to 0.36 µg g−1 cheese, were generally higher than the iDNA, indicating that autolytic phenomena prevailed over intact cells after 8–12 months of ripening. In four of the seven cheeses, the same LAB species were detected in the eDNA and iDNA fractions by length-heterogeneity PCR, while in the remaining three samples, a higher number of species was highlighted in the eDNA compared to the corresponding iDNA. The sequential extraction of eDNA and iDNA can be applied to obtain additional information on the composition of the bacterial community in long-aged cheeses.

scholarly journals Selection of raw cow’s milk thermophilic lactic acid bacteria obtained from southwest Parana, Brazil, with potential use as autochthonous starter

2020 ◽  
Vol 23 ◽  
Author(s):  
Simone Beux ◽  
Carla Todescatto ◽  
João Francisco Marchi ◽  
Edimir Andrade Pereira
Keyword(s):  
Starter Culture ◽  
Lactobacillus Rhamnosus ◽  
Streptococcus Thermophilus ◽  
Lactobacillus Helveticus ◽  
Lactobacillus Fermentum ◽  
Cow’S Milk ◽  
Lactobacillus Species ◽  
Cow's Milk ◽  
Specific Pcr ◽  
Raw Cow’S Milk

Abstract This work aimed to isolate and identify Streptococcus and Lactobacillus species from raw cow’s milk obtained from Southwest Paraná - Brazil. We used randomly amplified polymorphic DNA (RAPD)-PCR to identify and type 58Streptococcus and 48 Lactobacillus isolates, of which 04 Streptococcus thermophilus and 02 Streptpcoccus macedonicus were confirmed by species-specific PCR and by sequencing of the 16S ribosomal RNA of 02Streptococcus lutetiensis/infantarius, 10 Lactobacillus fermentum, 03 Lactobacillus delbrueickii subspecies bulgaricus, 01 Lactobacillus rhamnosus/casei and 02 Lactobacillus helveticus. The results indicated predominance of Streptococcus thermophillus and Lactobacillus fermentum. Streptococcus thermophilus and Lactobacillus delbrueickii subspecies bulgaricus strains were tested on the basis of their acidification kinetics. Considerable variation between the Streptococcus thermophilus was observed for the maximum rate of acidification (Vm), with a maximum of -4.5 and minimum of -4.2 pH milliunits min-1. The Lactobacillus delbrueickii subspecies bulgaricus showed values between -8.4 and -7.1 pH milliunits min-1. These results suggest that strains characterized as having a high acidifying capacity, could be used as starters in cheesemaking. The ferments presented an excellent performance in the acidification process, generating adequate curves, characteristics of a starter culture.

Adopting traditional fermented foods as carriers for probiotics

2019 ◽  
Vol 50 (5) ◽  
pp. 841-852
Author(s):  
Ivan Muzira Mukisa ◽  
Stellah Byakika ◽  
Rehema Meeme ◽  
Alex Paul Wacoo ◽  
Wilbert Sybesma ◽  
...  
Keyword(s):  
Lactobacillus Rhamnosus ◽  
Streptococcus Thermophilus ◽  
Soluble Solids ◽  
Consumer Acceptability ◽  
Fermented Foods ◽  
Content Type ◽  
Microbial Counts ◽  
Overall Acceptability ◽  
Novel Approach ◽  
Traditional Fermented Foods

Purpose Traditional fermented products can be adopted as probiotic carriers. This study was aimed at evaluating the potential of using Obushera, a traditional sorghum beverage from Uganda, as a carrier for Lactobacillus rhamnosus yoba. Design/methodology/approach Probiotic Obushera was produced by fermenting sorghum malt with Lb. rhamnosus yoba 2012 and Streptococcus thermophilus C106 at 30 °C and at room temperature (21°C-25 °C) for 24 h. Acidity, pH, total soluble solids and microbial counts were monitored. Consumer acceptability and purchase index of probiotic Obushera were compared to four commercial non-probiotic brands. Shelf stability of probiotic Obushera was determined by monitoring changes in pH, acidity, soluble solids, microbial counts and consumer acceptability during refrigerated storage. Findings Lactobacillus rhamnosus yoba 2012 multiplied and lowered the pH of Obushera from 5.3 to < 4.0 (p < 0.0001) whilst increasing acidity from 0.21 to 0.46 per cent (p < 0.0001) in 9 h at 30 °C. Consumer acceptability varied with Obushera brand (p < 0.0001). The overall acceptability score of probiotic Obushera (score of 6.4 = like slightly) was similar to that of the two most acceptable commercial brands (scores of 5.8 and 6.6). Acidity, pH and Lb. rhamnosus counts of probiotic Obushera varied within 0.6 per cent –1.05 per cent (p < 0.0001), 3.3–3.4 (p < 0.0001), and 8.2-9.2 log cfu/ml (p < 0.0001), respectively during two months of storage. The overall acceptability of probiotic Obushera (scores of 6.9-7.8) did not change significantly during storage (p = 0.185). Practical Implications Traditional fermented foods such as Obushera can be adopted as carriers of probiotic microorganisms. Originality/value Use of commercial probiotic strains in traditional fermented foods is a novel approach that can be adopted to improve safety of traditional fermentations and health of consumers.

scholarly journals Functional prediction of environmental variables using metabolic networks

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Adèle Weber Zendrera ◽  
Nataliya Sokolovska ◽  
Hédi A. Soula
Keyword(s):  
Machine Learning ◽  
Growth Temperature ◽  
Environmental Variables ◽  
Metabolic Networks ◽  
Machine Learning Techniques ◽  
Underlying Structure ◽  
Glutathione Biosynthesis ◽  
Additional Information ◽  
Cold Environments ◽  
Novel Approach

AbstractIn this manuscript, we propose a novel approach to assess relationships between environment and metabolic networks. We used a comprehensive dataset of more than 5000 prokaryotic species from which we derived the metabolic networks. We compute the scope from the reconstructed graphs, which is the set of all metabolites and reactions that can potentially be synthesized when provided with external metabolites. We show using machine learning techniques that the scope is an excellent predictor of taxonomic and environmental variables, namely growth temperature, oxygen tolerance, and habitat. In the literature, metabolites and pathways are rarely used to discriminate species. We make use of the scope underlying structure—metabolites and pathways—to construct the predictive models, giving additional information on the important metabolic pathways needed to discriminate the species, which is often absent in other metabolic network properties. For example, in the particular case of growth temperature, glutathione biosynthesis pathways are specific to species growing in cold environments, whereas tungsten metabolism is specific to species in warm environments, as was hinted in current literature. From a machine learning perspective, the scope is able to reduce the dimension of our data, and can thus be considered as an interpretable graph embedding.

scholarly journals Стан мнестичних процесів і рівень білків клітинної адгезії (NCAM) у гіпокампі щурів за умов введення S-аденозил-L-метіоніну та пре/пробіотиків на тлі медикаментозно-індукованого ураження печінки

2019 ◽  
Vol 13 (3) ◽  
pp. 166-174
Author(s):  
В.І. Жилюк ◽  
Г.О. Ушакова ◽  
Ю.В. Харченко ◽  
Д. В. Муравйова
Keyword(s):  
Lactobacillus Acidophilus ◽  
Lactobacillus Rhamnosus ◽  
Streptococcus Thermophilus ◽  
Lactobacillus Delbrueckii ◽  
Lactobacillus Delbrueckii Subsp

Мета дослідження – вивчення впливу S-аденозил-L-метіоніну, пре/пробіотиків та їхнього сумісного застосування на мнестичні процеси та рівень білків клітинної адгезії (NCAM) у гіпокампі щурів за умов тривалого введення рифампіцину та ізоніазиду. Дослідження проводили на 68 білих щурах-самцях лінії Wistar масою 180–220 г. Експериментальну модель токсичного медикаментозно-індукованого ураження печінки (МІУП) відтворювали шляхом повторних внутрішньошлункових (в/ш) введень ізоніазиду та рифампіцину у дозах 50 мг/кг і 86 мг/кг маси тіла відповідно протягом 28 діб. Тварин було розподілено на п’ять груп (n = 8 у кожній). І – інтактна, ІІ – контроль (МІУП). Щурам ІІІ групи протягом останніх 14 діб експерименту за 1 год до введення туберкулостатиків внутрішньом’язово вводили S-аденозил-L-метіонін у дозі 35 мг/кг. Щури IV групи в/ш отримували комбіновану терапію, що поєднувала пребіотик Лактулозу в дозі 2680 мг/кг і пробіотик, що містить 4 млрд активних клітин (КУО): Lactobacillus acidophilus, Lactobacillus rhamnosus, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. Bulgaricus у дозі 1 КУО/кг. Тваринам V групи проводили потрійну комбіновану фармакотерапію з усіма препаратами, які застосовували в ІІІ і ІV групах у відповідних дозах. Ноотропну активність оцінювали в тесті умовної реакції пасивного уникнення (УРПУ) за умов введення скополаміну. Для оцінки впливу скополаміну на процеси навчання використані значення показників груп тварин (інтактні, n = 10; МІУП, n = 18), які попередньо не отримували цей препарат. Кількісний уміст NCAM визначали за допомогою інгібіторного методу імуноферментного аналізу з використанням моноспецифічних антитіл щодо NCAM (Аbcam, Англія) і відповідного очищеного білка як стандарту (Аbnovo, США) у цитозольній фракції гомогенату гіпокампа. Отриманий цифровий матеріал обробляли методом варіаційної статистики за допомогою програми статистичного аналізу StatPlus, AnalystSoft Inc. Версія 6 (http://www.analystsoft.com/ru/). Результати експериментів свідчать про те, що курсове введення S-аденозил-L-метіоніну покращувало процеси навчання за умов введення скополаміну, що проявлялося суттєвим збільшенням латентного періоду в 3,14 разу (Р = 0,027) порівняно з групою тварин з МІУП, а число тварин з набутою навичкою складало 62,5 % (Р = 0,007). При цьому за умов введення цього препарату спостерігалося зростання на 38,7 % (Р = 0,004) рівнів цитоплазматичної форми NCAM у гіпокампі, яке ймовірно мало компенсаторний характер. Характерно, що корекція стану мікробіоти кишечника також може чинити позитивний вплив на мнестичні функції за умов тривалого введення туберкулостатиків, однак за своїм ноотропним потенціалом комбінація Лактулоза/Йогурт поступалася S-аденозил-Lметіоніну, хоча рівні NCAM у гіпокампі зростали на 39,6 % (Р = 0,004) порівняно з групою тварин з МІУП. Слід зазначити, що сумісне застосування пребіотика та пробіотика з S-аденозил-L-метіоніном не супроводжувалося підвищенням ноотропної активності. Отримані дані експериментально обґрунтовують застосування насамперед S-аденозил-Lметіоніну, а також пре/пробіотиків як перспективних засобів корекції когнітивних порушень за медикаментозних гепатитів. Необхідні подальші дослідження щодо можливості комбінованого використання зазначених засобів за умов вказаної патології.

scholarly journals Massive sequencing of artisan cheeses from raw sheep’s milk

2019 ◽  
Vol 6 (5) ◽  
pp. 236-240
Author(s):  
Ruiz Julia ◽  
Etcheverria Analia ◽  
Padola Nora ◽  
Kruger Alejandra ◽  
Medina Luis
Keyword(s):  
Dairy Products ◽  
Food Industry ◽  
Streptococcus Thermophilus ◽  
Molecular Techniques ◽  
Native Flora ◽  
Three Samples ◽  
Massive Sequencing ◽  
Traditional Fermentation ◽  
Antibacterial Capacity

Lactic acid bacteria (LAB) are used in the food industry to confer aromatic characteristics and their antibacterial capacity. In this study the native flora of LAB that participates in the traditional fermentation of semi-hard cheeses made with raw sheep's milk from the region of Andalusia, Spain was analyzed. Three samples of four different commercial cheeses were taken. Massive sequencing was carried out to identify the lactic and accompanying flora. Predominant lactic flora was Lactococcus lactis, Streptococcus thermophilus, Lactobacillus paracasei and Lactococcus raffinolactis, and to a lesser extent other species of the genera Lactobacillus, Streptococcus, Pediococcus and Leuconestoc. The accompanying flora was composed of species of the genera Mycoplasma, Pseudomonas, Acinetobacter, Chryseobacterium, Mannheimia, Trueperella, Enterococcus, Vibrio, Serratia, Macrococcus, Staphylococcus, Massilia, Flavobacterium, Yersinia, Gallaecimonas, Hafnia, Leclercia, Obesumbacterium, Morganella and Kluyvera. These results show that modern molecular techniques are very good tools to identify natural LABs of artisanal dairy products. The characterization of the native flora of the artisanal cheese allows us to evaluate the microbiological diversity of the natural population of LAB and the symbiosis with another type of flora.

scholarly journals Measuring Vascular Recovery Rate After Exercise

Proceedings ◽  
2018 ◽  
Vol 4 (1) ◽  
pp. 12 ◽  
Author(s):  
Halil Dijab ◽  
Jordi Alastruey ◽  
Peter Charlton
Keyword(s):  
Heart Rate ◽  
Recovery Rate ◽  
Wearable Sensors ◽  
Cardiovascular Fitness ◽  
Wave Shape ◽  
Future Studies ◽  
Additional Information ◽  
Novel Approach ◽  
Pulse Waves ◽  
Future Work

The rate at which an individual recovers from exercise is known to be indicative of cardiovascular risk. It has been widely shown that the reduction in heart rate immediately after exercise is predictive of mortality. However, little research has been conducted into whether the time taken for the blood vessels to return to normal is also indicative of risk. In this study, we present a novel approach to assess vascular recovery rate (VRR) using the photoplethysmogram (PPG) signal, which is monitored by smart wearables. The VORTAL dataset (http://peterhcharlton.github.io/RRest/) was used for this study, containing PPG signals from 39 healthy subjects before (baseline) and after exercise. 31 VRR indices were extracted from the PPG pulse wave shape, as well as heart rate for comparison. The rate at which indices returned to baseline after exercise was quantified, and the consistency of changes between subjects was assessed statistically. Many VRR indices exhibited changes after exercise which were consistent between subjects. Indices derived from the timings and second derivative of pulse waves were identified as candidates for future work. The rate at which the indices returned to baseline differed between indices and subjects, indicating that they may provide additional information beyond that of heart rate, and that they may be useful for stratifying subjects. This study demonstrated the feasibility of assessing VRR after exercise from the PPG. Future studies should investigate whether VRR indices are associated with cardiovascular fitness, and the potential utility of incorporating the indices into wearable sensors.

Effect of the addition of extracts of thermophilic lactobacilli on acid production byStreptococcus thermophilusin milk

1981 ◽  
Vol 48 (1) ◽  
pp. 139-148 ◽  
Author(s):  
Denis H. Hemme ◽  
Véronique Schmal ◽  
Jean E. Auclair
Keyword(s):  
Acid Production ◽  
Streptococcus Thermophilus ◽  
Lactobacillus Helveticus ◽  
Caseinolytic Activity ◽  
Hard Cheese

SummarySoluble extracts of 20 strains of thermophilic lactobacilli (Lactobacillus helveticus, L.lactisand L.bulgaricus) were prepared and added to milk for the culture of 10 strains ofStreptococcus thermophilus. Acid production was stimulated in 64·5% of cases for 9 of these 10 strains. The L.helveticusextracts were the most stimulatory, but the same extracts did not always strongly stimulate each strain ofStr. thermophilus. The stimulatory effects observed varied with the volume of extract and the strain ofStr. thermophilus. The exception wasStr. thermophilus385, which was never stimulated. The stimulatory effects observed were due to aminopeptidases present in the lactobacillus extracts and were not related to a general caseinolytic activity. The possible addition of such extracts to milk for cooked hard cheese is discussed.

Influence of thermophilic lactic acid bacteria strains on propionibacteria growth and lactate consumption in an Emmental juice-like medium

1999 ◽  
Vol 66 (1) ◽  
pp. 105-113 ◽  
Author(s):  
ANNE THIERRY ◽  
DELPHINE SALVAT-BRUNAUD ◽  
JEAN-LOUIS MAUBOIS
Keyword(s):  
Amino Acids ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Free Amino Acids ◽  
Free Amino ◽  
Streptococcus Thermophilus ◽  
Lactobacillus Helveticus ◽  
The Media ◽  
Lactate Consumption ◽  
Thermophilic Lactic Acid Bacteria

Swiss-type cheeses such as Emmental are characterized by the successive development of thermophilic lactic acid bacteria (TLAB) and propionibacteria. The aim of this study was to determine whether the choice of TLAB strain influenced propionibacteria. TLAB and propionibacteria were cultured sequentially under the conditions prevailing in cheese. Firstly, 11 Emmental juice-like media were prepared by fermenting casein-enriched milk with pure or mixed cultures of TLAB (Lactobacillus helveticus, Lb. delbrueckii subsp. lactis and Streptococcus thermophilus), differing in their proteolytic activities. TLAB cells were then removed by microfiltration. Finally, five strains of Propionibacterium freudenreichii were grown on these media at 24°C under anaerobiosis and their growth characteristics and lactate consumption determined. The media mainly differed in their contents of peptides (1·9–5·3 g/kg) and free amino acids (1·0–5·6 g/kg) and the proportions of lactate isomers (42–92% of the L(+) isomer). Propionibacteria were significantly (P<0·05) influenced by TLAB strains (differences in doubling times of up to 20% and differences in lactate consumption after 600 h culture of up to 52%). The influence of TLAB was similar for all the propionibacteria tested, depended on the TLAB strains and could not be generalized to the TLAB species. Propionibacteria were stimulated by high peptide levels, low levels of free amino acids and NaCl, a low proportion of L(+)-lactate and other undetermined factors. However, variations due to TLAB were less than those between propionibacteria strains.

Cloning and Characterization of a Prolinase Gene (pepR) from Lactobacillus rhamnosus

1998 ◽  
Vol 64 (5) ◽  
pp. 1831-1836 ◽  
Author(s):  
Pekka Varmanen ◽  
Terhi Rantanen ◽  
Airi Palva ◽  
Soile Tynkkynen
Keyword(s):  
Lactobacillus Rhamnosus ◽  
Gene Replacement ◽  
Lactobacillus Helveticus ◽  
Open Reading Frame ◽  
Transcriptional Unit ◽  
Wild Type ◽  
Reading Frame ◽  
Gene Expressing ◽  
Produce Acid

ABSTRACT A peptidase gene expressingl-proline-β-naphthylamide-hydrolyzing activity was cloned from a gene library of Lactobacillus rhamnosus 1/6 isolated from cheese. Peptidase-expressing activity was localized in a 1.5-kbSacI fragment. A sequence analysis of the SacI fragment revealed the presence of one complete open reading frame (ORF1) that was 903 nucleotides long. The ORF1-encoded 34.2-kDa protein exhibited 68% identity with the PepR protein from Lactobacillus helveticus. Additional sequencing revealed the presence of another open reading frame (ORF2) following pepR; this open reading frame was 459 bp long. Northern (RNA) and primer extension analyses indicated that pepR is expressed both as a monocistronic transcriptional unit and as a dicistronic transcriptional unit with ORF2. Gene replacement was used to construct a PepR-negative strain of L. rhamnosus. PepR was shown to be the primary enzyme capable of hydrolyzing Pro-Leu in L. rhamnosus. However, the PepR-negative mutant did not differ from the wild type in its ability to grow and produce acid in milk. The clonedpepR expressed activity against dipeptides with N-terminal proline residues. Also, Met-Ala, Leu-Leu, and Leu-Gly-Gly and the chromogenic substrates l-leucine-β-naphthylamide andl-phenylalanine-β-naphthylamide were hydrolyzed by the PepR of L. rhamnosus.

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