scholarly journals A Versatile Liquid Culture Method to Control the <i>in Vitro</i> Development of Shoot and Root Apical Meristems of Bamboo Plants

2020 ◽  
Vol 11 (02) ◽  
pp. 262-275
Author(s):  
Most Tanziman Ara ◽  
Taiji Nomura ◽  
Yasuo Kato ◽  
Shinjiro Ogita
Keyword(s):  
Liquid Culture ◽  
Culture Method ◽  
Apical Meristems

scholarly journals Enhancement of Campylobacter hepaticus culturing to facilitate downstream applications

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Canh Phung ◽  
Timothy B. Wilson ◽  
José A. Quinteros ◽  
Peter C. Scott ◽  
Robert J. Moore ◽  
...  
Keyword(s):  
Liquid Culture ◽  
Vaccine Development ◽  
Genomic Analysis ◽  
Bacterial Biomass ◽  
In Silico Analysis ◽  
Culture Method ◽  
Efficient Production ◽  
Culture Density ◽  
Brucella Broth

AbstractCampylobacter hepaticus causes Spotty Liver Disease (SLD) in chickens. C. hepaticus is fastidious and slow-growing, presenting difficulties when growing this bacterium for the preparation of bacterin vaccines and experimental disease challenge trials. This study applied genomic analysis and in vitro experiments to develop an enhanced C. hepaticus liquid culture method. In silico analysis of the anabolic pathways encoded by C. hepaticus revealed that the bacterium is unable to biosynthesise l-cysteine, l-lysine and l-arginine. It was found that l-cysteine added to Brucella broth, significantly enhanced the growth of C. hepaticus, but l-lysine or l-arginine addition did not enhance growth. Brucella broth supplemented with l-cysteine (0.4 mM), l-glutamine (4 mM), and sodium pyruvate (10 mM) gave high-density growth of C. hepaticus and resulted in an almost tenfold increase in culture density compared to the growth in Brucella broth alone (log10 = 9.3 vs 8.4 CFU/mL). The type of culture flask used also significantly affected C. hepaticus culture density. An SLD challenge trial demonstrated that C. hepaticus grown in the enhanced culture conditions retained full virulence. The enhanced liquid culture method developed in this study enables the efficient production of bacterial biomass and therefore facilitates further studies of SLD biology and vaccine development.

TWO MACHINES FOR IN VITRO PROPAGATION OF PLANTS IN LIQUID MEDIA

1983 ◽  
Vol 63 (1) ◽  
pp. 311-316 ◽  
Author(s):  
ROBERT E. HARRIS ◽  
EDWIN B. B. MASON
Keyword(s):  
Nicotiana Tabacum ◽  
In Vitro Propagation ◽  
Liquid Culture ◽  
Culture Method ◽  
The Other ◽  
Liquid Media ◽  
Wide Mouth ◽  
Agar Media ◽  
Two Machines

Two machines are described, one to tilt 400 50-mL or 320 125-mL Erlenmeyer flasks, and the other to rock 120 455-mL or 70 910-mL square wide-mouth Mason jars, or combinations of the different container sizes. In grapes, explants transferred to liquid media on the machines after 28 days on agar produced seven times as many shoots in 90 days as explants maintained on agar media. Similar increases were obtained with Arctostaphylos uva ursi, Fuchsia hybrida ’Swingtime,’ Amelanchier alnifolia, and Nicotiana tabacum ’Xanthi-nc.’ The liquid culture method also reduces cost by using less media and agar.Key words: Machines, in vitro, propagation, liquid-media

scholarly journals A Rapid Method for Quantifying Viable Mycobacterium avium subsp. paratuberculosis in Cellular Infection Assays

2016 ◽  
Vol 82 (18) ◽  
pp. 5553-5562 ◽  
Author(s):  
Hannah B. Pooley ◽  
Kumudika de Silva ◽  
Auriol C. Purdie ◽  
Douglas J. Begg ◽  
Richard J. Whittington ◽  
...  
Keyword(s):  
Growth Rate ◽  
Quantitative Pcr ◽  
Mycobacterium Avium ◽  
Large Scale ◽  
Liquid Culture ◽  
Vaccine Development ◽  
Culture Method ◽  
Content Type ◽  
Antimicrobial Screening

ABSTRACTDetermining the viability of bacteria is a key outcome ofin vitrocellular infection assays. Currently, this is done by culture, which is problematic for fastidious slow-growing bacteria such asMycobacterium aviumsubsp.paratuberculosis, where it can take up to 4 months to confirm growth. This study aimed to identify an assay that can rapidly quantify the number of viableM. aviumsubsp.paratuberculosiscells in a cellular sample. Three commercially available bacterial viability assays along with a modified liquid culture method coupled with high-throughput quantitative PCR growth detection were assessed. Criteria for assessment included the ability of each assay to differentiate live and deadM. aviumsubsp.paratuberculosisorganisms and their accuracy at low bacterial concentrations. Using the culture-based method,M. aviumsubsp.paratuberculosisgrowth was reliably detected and quantified within 2 weeks. There was a strong linear association between the 2-week growth rate and the initial inoculum concentration. The number of viableM. aviumsubsp.paratuberculosiscells in an unknown sample was quantified based on the growth rate, by using growth standards. In contrast, none of the commercially available viability assays were suitable for use with samples fromin vitrocellular infection assays.IMPORTANCERapid quantification of the viability ofMycobacterium aviumsubsp.paratuberculosisin samples fromin vitrocellular infection assays is important, as it allows these assays to be carried out on a large scale.In vitrocellular infection assays can function as a preliminary screening tool, for vaccine development or antimicrobial screening, and also to extend findings derived from experimental animal trials. Currently, by using culture, it takes up to 4 months to obtain quantifiable results regardingM. aviumsubsp.paratuberculosisviability after anin vitroinfection assay; however, with the quantitative PCR and liquid culture method developed, reliable results can be obtained at 2 weeks. This method will be important for vaccine and antimicrobial screening work, as it will allow a greater number of candidates to be screened in the same amount of time, which will increase the likelihood that a favorable candidate will be found to be subjected to further testing.

scholarly journals Isolation of Mycobacterium avium Subsp. Paratuberculosis in the Feces and Tissue of Small Ruminants Using a Non-Automated Liquid Culture Method

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 20
Author(s):  
Luigi De Grossi ◽  
Davide Santori ◽  
Antonino Barone ◽  
Silvia Abbruzzese ◽  
Matteo Ricchi ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Liquid Culture ◽  
Culture Media ◽  
Small Ruminants ◽  
Culture Method ◽  
Mycobacterium Avium Subsp ◽  
Type I ◽  
Culture Methods ◽  
Specific Pcr ◽  
Mycobacterium Avium Subsp Paratuberculosis

Paratuberculosis is a chronic disease of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Since isolation of MAP type I (S) is rarely reported in Italy, our research was aimed at isolating, by an inexpensive liquid culture manual method, this type of MAP isolates. At first, we used an ELISA to point out to serologically positive samples from five flocks. Secondly, we used a fecal direct IS900-qPCR on the ELISA positive samples, in order to detect shedder animals. Feces from IS900-qPCR positive samples were inoculated in solid and liquid culture media. IS900-qPCR was further used to test the growth of MAP isolates in liquid medium, which were further confirmed by f57-qPCR and submitted to typing by specific PCR in order to identify the MAP type. Twenty-eight samples (24 fecal and four tissutal samples) were processed by culture methods, resulting in the isolation of six type I MAP field isolates. Notably, no isolates were recovered by solid media, underlining the utility of this liquid method. Few data about this type of MAP are currently available in Italy, and further analyses should be carried out in order to study the origin and epidemiology of type I strains circulating in Italy.

scholarly journals The Diversity of Root-Associated Endophytic Fungi from Four Epiphytic Orchids in China

Diversity ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 197
Author(s):  
Tao Wang ◽  
Miao Chi ◽  
Ling Guo ◽  
Donghuan Liu ◽  
Yu Yang ◽  
...  
Keyword(s):  
Endophytic Fungi ◽  
Southern China ◽  
Culture Method ◽  
Amplicon Sequencing ◽  
Community Diversity ◽  
Mycorrhizal Symbiosis ◽  
Epiphytic Orchids ◽  
Culture Independent ◽  
Almost All

Root-associated endophytic fungi (RAF) are found asymptomatically in almost all plant groups. However, little is known about the compositions and potential functions of RAF communities associated with most Orchidaceae species. In this study, the diversity of RAF was examined in four wild epiphytic orchids, Acampe rigida, Doritis pulcherrima, Renanthera coccinea, and Robiquetia succisa, that occur in southern China. A culture-independent method involving Illumina amplicon sequencing, and an in vitro culture method, were used to identify culturable fungi. The RAF community diversity differed among the orchid roots, and some fungal taxa were clearly concentrated in a certain orchid species, with more OTUs being detected. By investigating mycorrhizal associations, the results showed that 28 (about 0.8%) of the 3527 operational taxonomic units (OTUs) could be assigned as OMF, while the OTUs of non-mycorrhizal fungal were about 99.2%. Among the OMFs, Ceratobasidiaceae OTUs were the most abundant with different richness, followed by Thelephoraceae. In addition, five Ceratobasidium sp. strains were isolated from D. pulcherrima, R. succisa, and R. coccinea roots with high separation rates. These culturable Ceratobasidium strains will provide materials for host orchid conservation and for studying the mechanisms underlying mycorrhizal symbiosis.

scholarly journals Mesenchymal Stem/Stromal Cells from Discarded Neonatal Sternal Tissue: In Vitro Characterization and Angiogenic Properties

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Shuyun Wang ◽  
Lakshmi Mundada ◽  
Eric Colomb ◽  
Richard G. Ohye ◽  
Ming-Sing Si
Keyword(s):  
Bone Marrow ◽  
Cardiac Surgery ◽  
Stromal Cells ◽  
Umbilical Vein ◽  
Culture Method ◽  
Tissue Perfusion ◽  
Explant Culture ◽  
Critical Congenital Heart Disease ◽  
Surface Marker Expression

Autologous and nonautologous bone marrow mesenchymal stem/stromal cells (MSCs) are being evaluated as proangiogenic agents for ischemic and vascular disease in adults but not in children. A significant number of newborns and infants with critical congenital heart disease who undergo cardiac surgery already have or are at risk of developing conditions related to inadequate tissue perfusion. During neonatal cardiac surgery, a small amount of sternal tissue is usually discarded. Here we demonstrate that MSCs can be isolated from human neonatal sternal tissue using a nonenzymatic explant culture method. Neonatal sternal bone MSCs (sbMSCs) were clonogenic, had a surface marker expression profile that was characteristic of bone marrow MSCs, were multipotent, and expressed pluripotency-related genes at low levels. Neonatal sbMSCs also demonstrated in vitro proangiogenic properties. Sternal bone MSCs cooperated with human umbilical vein endothelial cells (HUVECs) to form 3D networks and tubes in vitro. Conditioned media from sbMSCs cultured in hypoxia also promoted HUVEC survival and migration. Given the neonatal source, ease of isolation, and proangiogenic properties, sbMSCs may have relevance to therapeutic applications.

scholarly journals In vitro mouse spermatogenesis with an organ culture method in chemically defined medium

PLoS ONE ◽  
2018 ◽  
Vol 13 (2) ◽  
pp. e0192884 ◽  
Author(s):  
Hiroyuki Sanjo ◽  
Mitsuru Komeya ◽  
Takuya Sato ◽  
Takeru Abe ◽  
Kumiko Katagiri ◽  
...  
Keyword(s):  
Organ Culture ◽  
Culture Method ◽  
Defined Medium ◽  
Chemically Defined Medium
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