scholarly journals Evolutionary changes of an intestinalLactobacillus reuteriduring probiotic manufacture

2019 ◽  
Author(s):  
Lázaro López ◽  
Paúl Cardenas ◽  
María B. Prado ◽  
Gabriel Trueba
Keyword(s):  
Microbial Community ◽  
Culture Medium ◽  
Lactobacillus Reuteri ◽  
Culture Media ◽  
Probiotic Bacteria ◽  
Synonymous Mutations ◽  
Metabolic Genes ◽  
Evolutionary Changes ◽  
Mammalian Intestine ◽  
Improve Health

ABSTRACTProbiotic bacteria are frequently used to treat intestinal (and other types of diseases) or to improve health, however little is known about the evolutionary changes of these bacteria during probiotic manufacture. It has been observed that when bacteria adapt to a new environment, they lose the skills to thrive in the original niche. In this study, a strain ofLactobacillus reuteriwas isolated from the duodenum of a mouse and subjected to 150 serial passes in milk to simulate industrial propagation of probiotic bacteria. The strain adapted to milk outperformed its ancestor in milk, but it showed reduced aptitude to grow culture media, and possibly in mouse intestines. Bacterial adaptation to milk seemed to select a number of non-synonymous mutations in metabolic genes.IMPORTANCEBacteria in the microbiota must endure hostile conditions including antagonism from other members of the microbial community. Little is known about the evolution of probiotic bacteria in the dairy industry. We show that repeated growth ofL. reuteriin milk results in reduced ability to grow in culture medium and possibly colonize the mammalian intestine. Our results contribute to the notion that probiotic bacteria colonize only transiently intestines.

scholarly journals Cholesterol Assimilation byLactobacillusProbiotic Bacteria: AnIn VitroInvestigation

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Catherine Tomaro-Duchesneau ◽  
Mitchell L. Jones ◽  
Divya Shah ◽  
Poonam Jain ◽  
Shyamali Saha ◽  
...  
Keyword(s):  
Lactobacillus Reuteri ◽  
Culture Media ◽  
Lactobacillus Rhamnosus ◽  
Probiotic Bacteria ◽  
Bile Salt Hydrolase ◽  
Dietary Interventions ◽  
Cholesterol Levels ◽  
Probiotic Strains ◽  
Therapeutic Measures ◽  
Pharmaceutical Agents

Excess cholesterol is associated with cardiovascular diseases (CVD), an important cause of mortality worldwide. Current CVD therapeutic measures, lifestyle and dietary interventions, and pharmaceutical agents for regulating cholesterol levels are inadequate. Probiotic bacteria have demonstrated potential to lower cholesterol levels by different mechanisms, including bile salt hydrolase activity, production of compounds that inhibit enzymes such as 3-hydroxy-3-methylglutaryl coenzyme A, and cholesterol assimilation. This work investigates 11Lactobacillusstrains for cholesterol assimilation. Probiotic strains for investigation were selected from the literature:Lactobacillus reuteriNCIMB 11951,L. reuteriNCIMB 701359,L. reuteriNCIMB 702655,L. reuteriNCIMB 701089,L. reuteriNCIMB 702656,Lactobacillus fermentumNCIMB 5221,L. fermentumNCIMB 8829,L. fermentumNCIMB 2797,Lactobacillus rhamnosusATCC 53103 GG,Lactobacillus acidophilusATCC 314, andLactobacillus plantarumATCC 14917. Cholesterol assimilation was investigated in culture media and under simulated intestinal conditions. The best cholesterol assimilator wasL. plantarumATCC 14917 (15.18 ± 0.55 mg/1010 cfu) in MRS broth.L. reuteriNCIMB 701089 assimilated over 67% (2254.70 ± 63.33 mg/1010 cfu) of cholesterol, the most of all the strains, under intestinal conditions. This work demonstrates that probiotic bacteria can assimilate cholesterol under intestinal conditions, withL. reuteriNCIMB 701089 showing great potential as a CVD therapeutic.

Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Water Concentration ◽  
Medium Composition ◽  
Coconut Water ◽  
Orchid Species ◽  
Randomized Design ◽  
Wet Weight ◽  
Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

scholarly journals Untargeted Metabolomics Approach for the Discovery of Environment-Related Pyran-2-ones Chemodiversity in a Marine-Sourced Penicillium restrictum

Marine Drugs ◽  
2021 ◽  
Vol 19 (7) ◽  
pp. 378
Author(s):  
Van-Tuyen Le ◽  
Samuel Bertrand ◽  
Thibaut Robiou du Pont ◽  
Fabrice Fleury ◽  
Nathalie Caroff ◽  
...  
Keyword(s):  
Culture Medium ◽  
Blue Mussel ◽  
Culture Media ◽  
Principal Component ◽  
Chemical Diversity ◽  
Untargeted Metabolomics ◽  
High Chemical ◽  
Medium Mass ◽  
Metabolomics Study ◽  
Penicillium Restrictum

Very little is known about chemical interactions between fungi and their mollusc host within marine environments. Here, we investigated the metabolome of a Penicillium restrictum MMS417 strain isolated from the blue mussel Mytilus edulis collected on the Loire estuary, France. Following the OSMAC approach with the use of 14 culture media, the effect of salinity and of a mussel-derived medium on the metabolic expression were analysed using HPLC-UV/DAD-HRMS/MS. An untargeted metabolomics study was performed using principal component analysis (PCA), orthogonal projection to latent structure discriminant analysis (O-PLSDA) and molecular networking (MN). It highlighted some compounds belonging to sterols, macrolides and pyran-2-ones, which were specifically induced in marine conditions. In particular, a high chemical diversity of pyran-2-ones was found to be related to the presence of mussel extract in the culture medium. Mass spectrometry (MS)- and UV-guided purification resulted in the isolation of five new natural fungal pyran-2-one derivatives—5,6-dihydro-6S-hydroxymethyl-4-methoxy-2H-pyran-2-one (1), (6S, 1’R, 2’S)-LL-P880β (3), 5,6-dihydro-4-methoxy-6S-(1’S, 2’S-dihydroxy pent-3’(E)-enyl)-2H-pyran-2-one (4), 4-methoxy-6-(1’R, 2’S-dihydroxy pent-3’(E)-enyl)-2H-pyran-2-one (6) and 4-methoxy-2H-pyran-2-one (7)—together with the known (6S, 1’S, 2’S)-LL-P880β (2), (1’R, 2’S)-LL-P880γ (5), 5,6-dihydro-4-methoxy-2H-pyran-2-one (8), (6S, 1’S, 2’R)-LL-P880β (9), (6S, 1’S)-pestalotin (10), 1’R-dehydropestalotin (11) and 6-pentyl-4-methoxy-2H-pyran-2-one (12) from the mussel-derived culture medium extract. The structures of 1-12 were determined by 1D- and 2D-MMR experiments as well as high-resolution tandem MS, ECD and DP4 calculations. Some of these compounds were evaluated for their cytotoxic, antibacterial, antileishmanial and in-silico PTP1B inhibitory activities. These results illustrate the utility in using host-derived media for the discovery of new natural products.

scholarly journals Effects of Lactobacillus reuteri and Streptomyces coelicolor on Growth Performance of Broiler Chickens

2021 ◽  
Vol 9 (6) ◽  
pp. 1341
Author(s):  
Sarayu Bhogoju ◽  
Collins N. Khwatenge ◽  
Thyneice Taylor-Bowden ◽  
Gabriel Akerele ◽  
Boniface M. Kimathi ◽  
...  
Keyword(s):  
Broiler Chickens ◽  
Lactobacillus Reuteri ◽  
Body Weight Gain ◽  
Streptomyces Coelicolor ◽  
Control Diet ◽  
Probiotic Bacteria ◽  
Feed Consumption ◽  
Feed Conversion ◽  
Wheat Middlings ◽  
Dietary Treatments

There are well documented complications associated with the continuous use of antibiotics in the poultry industry. Over the past few decades, probiotics have emerged as viable alternatives to antibiotics; however, most of these candidate probiotic microorganisms have not been fully evaluated for their effectiveness as potential probiotics for poultry. Recent evaluation of a metagenome of broiler chickens in our laboratory revealed a prevalence of Lactobacillus reuteri (L. reuteri) and Actinobacteria class of bacteria in their gastrointestinal tract. In this study Lactobacillus reuteri and Streptomyces coelicolor (S. coelicolor) were selected as probiotic bacteria, encapsulated, and added into broiler feed at a concentration of 100 mg/kg of feed. In an 8-week study, 240 one day-old chicks were randomly assigned to four dietary treatments. Three dietary treatments contained two probiotic bacteria in three different proportions (L. reuteri and S. coelicolor individually at 100 ppm, and mixture of L. reuteri and S. coelicolor at 50 ppm each). The fourth treatment had no probiotic bacteria and it functioned as the control diet. L. reuteri and S. coelicolor were added to the feed by using wheat middlings as a carrier at a concentration of 100 ppm (100 mg/kg). Chickens fed diets containing L. reuteri and S. coelicolor mixture showed 2% improvement in body weight gain, 7% decrease in feed consumption, and 6–7% decrease in feed conversion ratios. This research suggests that L. reuteri and S. coelicolor have the potential to constitute probiotics in chickens combined or separately, depending on the desired selection of performance index.

scholarly journals Rheological and Microstructural Features of Plant Culture Media Doped with Biopolymers: Influence on the Growth and Physiological Responses of In Vitro-Grown Shoots of Thymus lotocephalus

2021 ◽  
Vol 2 (2) ◽  
pp. 538-553
Author(s):  
Natacha Coelho ◽  
Alexandra Filipe ◽  
Bruno Medronho ◽  
Solange Magalhães ◽  
Carla Vitorino ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Average Pore Size ◽  
Physiological Responses ◽  
Gum Arabic ◽  
Shoot Elongation ◽  
Hydroxyethyl Cellulose ◽  
Plant Culture

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.

scholarly journals Influenza Virus Hemagglutinin and Neuraminidase, but Not the Matrix Protein, Are Required for Assembly and Budding of Plasmid-Derived Virus-Like Particles

2007 ◽  
Vol 81 (13) ◽  
pp. 7111-7123 ◽  
Author(s):  
Benjamin J. Chen ◽  
George P. Leser ◽  
Eiji Morita ◽  
Robert A. Lamb
Keyword(s):  
Influenza Virus ◽  
Culture Medium ◽  
Matrix Protein ◽  
Culture Media ◽  
Protein Composition ◽  
Multivesicular Body ◽  
Viral Proteins ◽  
Dominant Negative ◽  
The Matrix ◽  
Overexpression System

ABSTRACT For influenza virus, we developed an efficient, noncytotoxic, plasmid-based virus-like particle (VLP) system to reflect authentic virus particles. This system was characterized biochemically by analysis of VLP protein composition, morphologically by electron microscopy, and functionally with a VLP infectivity assay. The VLP system was used to address the identity of the minimal set of viral proteins required for budding. Combinations of viral proteins were expressed in cells, and the polypeptide composition of the particles released into the culture media was analyzed. Contrary to previous findings in which matrix (M1) protein was considered to be the driving force of budding because M1 was found to be released copiously into the culture medium when M1 was expressed by using the vaccinia virus T7 RNA polymerase-driven overexpression system, in our noncytotoxic VLP system M1 was not released efficiently into the culture medium. Additionally, hemagglutinin (HA), when treated with exogenous neuraminidase (NA) or coexpressed with viral NA, could be released from cells independently of M1. Incorporation of M1 into VLPs required HA expression, although when M1 was omitted from VLPs, particles with morphologies similar to those of wild-type VLPs or viruses were observed. Furthermore, when HA and NA cytoplasmic tail mutants were included in the VLPs, M1 failed to be efficiently incorporated into VLPs, consistent with a model in which the glycoproteins control virus budding by sorting to lipid raft microdomains and recruiting the internal viral core components. VLP formation also occurred independently of the function of Vps4 in the multivesicular body pathway, as dominant-negative Vps4 proteins failed to inhibit influenza VLP budding.

P-203 Applying artificial intelligence for ploidy prediction: The concentration of IL-6 in spent culture medium, blastocyst morphological grade and embryo morphokinetics as variables under consideration

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
B Aparicio Ruiz ◽  
L Bori ◽  
E Paya ◽  
M A Valera ◽  
A Quiñonero ◽  
...  
Keyword(s):  
Embryo Culture ◽  
Culture Medium ◽  
Culture Media ◽  
General Description ◽  
Ann Model ◽  
Non Invasive ◽  
Morphokinetic Parameters ◽  
Testing Dataset ◽  
Standard Deviations ◽  
The Impact

Abstract Study question Would it be possible to predict embryo ploidy by taking into account conventional morphological and morphokinetic parameters together with IL-6 concentration in spent culture medium? Summary answer Our artificial neural network (ANN) trained with blastocyst morphology, embryo morphokinetics and IL-6 concentration distinguished between euploid/aneuploid embryos in 65% of the testing dataset. What is known already The analysis of spent embryo culture media represents the protein and metabolic state of the embryo and could be a non-invasive method of obtaining information about embryo quality. The impact of the presence/absence of several proteins in embryo culture samples over clinical results has been widely studied. The IL-6 is one of the most mentioned protein for its effect on embryo development, implantation and likelihood of achieving a live birth. In this initial attempt, we examined the predictive value for euploidy of a model that took into account the concentration of IL-6 in the spent culture medium. Study design, size, duration This prospective study included 319 embryos with PGT-A results. Out of the total, 127 were euploid and 192 aneuploid embryos. Concentration of IL-6 in spent embryo culture media (collected on the day of trophectoderm biopsy-fifth/sixth day of development), morphokinetic parameters (division time to 2 cells-t2; to 3 cells-t3, to 4 cells-t4; to 5 cells-t5 and time of blastocyst formation-tB) and blastocyst morphological grade (according to ASEBIR criteria) were considered to predict the embryo ploidy. Participants/materials, setting, methods Embryos were cultured in EmbryoScope. The chromosome analysis was performed using next-generation sequence technology. The concentration of IL-6 was measured in 20µL of spent embryo culture media with ELISA kits. Morphokinetic parameters were automatically annotated and the blastocyst morphology was evaluated by senior embryologists based on blastocele expansion, inner cell mass and trophectoderm quality. All the embryos were divided into 70% for training, 15% for validating and 15% for testing our ANN model with MatLab®. Main results and the role of chance The general description for the euploid embryo population was the following: 2% of the embryos were graded as A, 71% were graded as B and 28% were graded as C; the means and standard deviations were 25.32±2.97 hours (h) for t2, 35.33±5.15h for t3, 37.30±5.43h for t4, 48.24±6.62h for t5 and 103.93±12.8h for tB; and the average of IL-6 concentration was 1.51±0.70 pg/ml. The general description for the aneuploid embryo population was the following: 1% of the embryos were graded as A, 48% were graded as B and 51% were graded as C; the means and standard deviations were 26.13±3.51h for t2, 36.70±4.29h for t3, 38.20±4.24h for t4, 49.86±6.89h for t5 and 107.10±8.29h for tB; and the average of IL-6 concentration was 1.47±0.71 pg/ml. Our ANN model showed a higher general success rate as we increased the variables considered in the final prediction of euploid embryos. The accuracy, sensitivity and specificity for the testing dataset were: 0.60, 0.12 and 0.87 with morphokinetic parameters; 0.63, 0.24 and 0.93 with morphokinetics and IL-6 concentration; and 0.65, 0.16 and 0.96 with morphokinetics, IL-6 concentration and blastocyst morphological grade. Limitations, reasons for caution The low sensitivity and high specificity achieved in our models indicated that they were more capable of detecting aneuploid than euploid embryos. As this was a preliminary study, the small number of embryos included in the test (n = 48) was also a limitation. Wider implications of the findings The results showed that our model tended to classify the embryos as aneuploid. More euploid embryos would be necessary to train our model and achieve better results in the prediction of chromosomally normal embryos. Further studies with large number of embryos and additional variables could improve the non-invasive ploidy prediction. Trial registration number not applicable

Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos

2021 ◽  
Author(s):  
Gabriela de Oliveira Fernandes ◽  
Marcella Pecora Milazzotto ◽  
Andrei Antonioni Guedes Fidelis ◽  
Taynan Stonoga Kawamoto ◽  
Ligiane de Oliveira Leme ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Biochemical Markers ◽  
Culture Medium ◽  
Culture Media ◽  
Ionization Mass ◽  
Metabolic Profiles ◽  
Bovine Embryos ◽  
The Media

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

scholarly journals STUDI BACILLUS FIRMUS SEBAGAI KANDIDAT PROBIOTIK DALAM MENGHADAPI Aeromonas hydrophila PADA MEDIA BUDIDAYA The Study of Bacillus firmus as Probiotic Candidate in Supressing Aeromonas hydrophila in Culture Media

2016 ◽  
Vol 11 (2) ◽  
pp. 111
Author(s):  
Alfabetian Harjuno Condro Haditomo ◽  
Angela Mariana Lusiastuti ◽  
Widanarni Widanarni
Keyword(s):  
Aeromonas Hydrophila ◽  
Pathogenic Bacteria ◽  
Culture Media ◽  
Inhibition Zone ◽  
Probiotic Bacteria ◽  
Bacterial Disease ◽  
Second Phase ◽  
Bacillus Firmus ◽  
Culture Methods

ABSTRAK   Pengendalian penyakit bakterial yang umum dilakukan dengan pemakaian antibiotik atau  bahan kimia sudah tidak diperbolehkan lagi karena menimbulkan patogen yang resisten  terhadap bahan kimia tersebut, terlebih jika penggunaan tidak sesuai dengan anjuran yang diberikan. Dampak negatif terhadap kesehatan konsumen berupa residu antibiotik juga menjadi pertimbangan yang harus diperhatikan. Manipulasi terhadap populasi mikroba yang berada di perairan guna pencegahan sebelum terjadinya serangan bakteri yang bersifat mematikan perlu dilakukan sebagaimana konsep probiotik sebagai biokontrol. Tujuan penelitian ini adalah menguji kandidat probiotik dalam menekan atau menghambat bakteri patogen Aeromonas hydrophila. Penelitian ini dilaksananakan dalam dua tahap. Tahap pertama adalah tahap pengujian bakteri kandidat probiotik secara in vitro menggunakan metode zona hambat dan kultur bersama pada media agar.  Tahap kedua adalah uji tentang bakteri kandidat probiotik dengan patogen pada media budidaya. Hasil terbaik penelitian tahap pertama pada  uji kultur bersama antara kandidat probiotik B. firmus dengan A. hydrophila pada skala in vitro adalah dengan penambahan probiotik  B. firmus sebanyak 108 cfu/ml. Sedangkan pada penelitian tahap kedua didapatkan hasil berturut-turut perlakuan D dengan tingkat kelangsungan hidup (SR) mencapai 90%, perlakuan C dengan SR 75%, perlakuan A dengan SR 50% dan perlakuan K dengan SR 50%.   Kata kunci: Bacillus firmus, probiotik, Aeromonas hydrophila, media budidaya   ABSTRACT  Controlling bacterial disease with the use of antibiotics or chemicals is no longer allowed as it results in pathogens that are resistant to the chemicals, especially when not in accordance with the recommendations provided. The negative impactsof the antibiotics residues on the consumers’ health  also need to be considered. Manipulation of microbial populations present in the waters as preventation before the lethal attack of bacteria needs to be done which is in accordance with the concept of probiotics as biocontrol.The purpose of this study was to test the probiotic candidates in suppressing or inhibiting pathogenic bacteria Aeromonas hydrophila. This study was conducted in two stages. The first stage was to test a candidate probiotic bacteria in vitro using culture methods and inhibition zone on the media together. The second stage wasto test candidate probiotic bacteria to pathogens on the cultivation media. The best results in the first phase of the research is shared culture test between probiotic candidate B. FIRMUS with A. hydrophila on vitro scale is the addition of the probiotic B. FIRMUS 108 cfu / ml. While in the second phase of the research results obtained successively: treatment D with a survival rate (SR) reaches 90%, treatment C with SR 75%, treatment A with SR 50% and treatment K with SR 50%. Keywords: Bacillus FIRMUS, probiotics, Aeromonas hydrophila, media cultivation

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