scholarly journals ON A METHOD OF ISOLATING AND IDENTIFYING BACILLUS TYPHOSUS, BASED ON A STUDY OF BACILLUS TYPHOSUS AND MEMBERS OF THE COLON GROUP IN SEMI-SOLID CULTURE MEDIA

1897 ◽  
Vol 2 (6) ◽  
pp. 677-700 ◽  
Author(s):  
Philip Hanson Hiss
Keyword(s):  
Typhoid Fever ◽  
Culture Media ◽  
Tap Water ◽  
Bacterial Species ◽  
Practical Application ◽  
Solid Culture ◽  
The Media ◽  
Semi Solid

Semi-solid culture media, and more especially media rendered semi-solid by temperatures of from 30° to 40° C., seem to have an important bearing in the differentiation of bacterial species, particularly those presenting various degrees of motility. In such media not only the effect of differences in consistence on the motility of an organism may be noted, but the effect produced by various chemicals and nutrient ingredients on the growth and motility may be readily observed. By systematically varying the constituents of such media it has been possible to produce a medium in which the behavior of Bacillus typhosus differentiates it from the various members of the colon group; and also to produce a medium in which the colonies of Bacillus typhosus assume a form which distinguishes them from the colonies of the colon bacilli in plate cultures. Bacillus typhosus alone of all the organisms investigated during these experiments has displayed both the power of giving rise to thread-forming colonies in the plating medium and that of the uniform clouding of the tube medium, hence these two characters may prove to be of great value in the identification of this organism. The practical application of the use of these media has led to the ready detection of Bacillus typhosus and its isolation from the stools of patients suffering from typhoid fever. No suspected water has been subjected to test, but from the investigation of artificially infected tap-water the media here described may be assumed to have an application in the detection of Bacillus typhosus in such waters.

scholarly journals Agar Desiccation – The Causes and How to Address Them

2021 ◽  
Author(s):  
Tammy Hassel
Keyword(s):  
Moisture Content ◽  
Environmental Monitoring ◽  
Culture Media ◽  
False Negative ◽  
Data Gathering ◽  
Petri Dish ◽  
Solid Culture ◽  
Critical Data ◽  
Negative Results ◽  
The Media

Solid culture media, or agar plates, are prone to desiccation. The combination of warm, dry air and limited moisture content to achieve a solid finish leads to inevitable drying out and desiccation. Cracks in the media, a loss of volume or shrinkage away from the sides of the Petri dish are evidence of desiccation. Overall, this reduces the capability of the media to support the growth of organisms. The reduction in growth support capability is particularly so if the microorganisms deposited on the surface of the media are stressed. Why is Desiccation an Issue It is a well-known fact that microbes have three basic requirements for growth; a food source, warmth and moisture. Reducing the moisture content risks inactivating and or inhibiting growth. The purpose of monitoring for microbes through the use of solid media is to capture microorganisms from the manufacturing environment and create favourable conditions to support their optimum growth, thus allowing them to replicate and form visible colonies for inspection. Viable environmental monitoring is a challenging process to optimise for; you are looking for the chance of capturing one of the very few microorganisms present in your extensive manufacturing facility. To do so, you use solid culture media to take samples, snapshots in time, of your environment. Firstly, you have a vast space to monitor, and your monitoring activity needs to find the very few microorganisms present. If you have set your monitoring programme up suitably and you successfully capture organism present in the environment but then add desiccation you risk losing critical data. Desiccation leads to a reduction in these favourable conditions and potentially a loss of critical data and false-negative results. Desiccation of media is considered a significant quality issue as it can lead to the inhibition of growth and or cell death. The action of drying out of the media can lead to the formation of a skin on the surface. The skin formation inhibits the recovery of organisms, through «bounce» or «air bounce». As the purpose of environmental monitoring is to capture and support the growth of contaminants, this issue will result in inconclusive data gathering. Desiccation is an inevitability. Cleanroom environments are by their very nature, high airflow facilities which, as you will see below, is a primary reason for the loss of moisture from agar plates. It is, therefore, important to ensure media qualification studies include a review that post use, your media, can meet the 70% growth rate recovery as specified in USP <1227>.

scholarly journals Fecundity of inbred fruit fly Drosophila melanogaster on different solid culture media: An analysis

2018 ◽  
Vol 10 (4) ◽  
pp. 1109-1114 ◽  
Author(s):  
Animesh Kumar Mohapatra ◽  
Priyamvada Pandey
Keyword(s):  
Drosophila Melanogaster ◽  
First Generation ◽  
Culture Media ◽  
Fruit Fly ◽  
Control Medium ◽  
Solid Culture ◽  
Pupal Development ◽  
Multiple Comparison Test ◽  
The Media ◽  
And Control

In the present study, wild-type Drosophila melanogaster collected from stock culture were sub-cultured in three different types of solid culture media  (corn, barley and wheat) and control medium for two weeks to produce F1 generation. The duration of larval and pupal development, number of pupal cases and hatched flies were scored for first generation. The results were analyzed by using one-way ANOVA, Bonferroni multiple comparison test and paired sample t-test. The control medium showed no pupal cases and hatched flies. Among all the three solid culture media tested, corn meal, barley meal and wheat meal, the latter showed highly significant results at p?0.001 than others. However, this parameter was not affected by the carbohydrate amount in the media. The present investigation is an attempt to evaluate the influence of different formulated solid culture media on the life span and reproduction of fruit flies.

scholarly journals APLIKASI SITOKININ TIPE PURIN DAN UREA PADA MULTIPLIKASI TUNAS ANIS (Pimpinellla anisum L.) IN VITRO

2020 ◽  
Vol 13 (1) ◽  
pp. 1
Author(s):  
OTIH ROSTIANA
Keyword(s):  
Essential Oils ◽  
Culture Media ◽  
Shoot Multiplication ◽  
Tropical Region ◽  
Herbaceous Plant ◽  
Solid Culture ◽  
Pimpinella Anisum ◽  
The Media ◽  
Initiation Stage

ABSTRAK<br />Anis (Pimpinella anisum L.) merupakan tanaman herba tahunan<br />yang termasuk ke dalam famili Umbelliferae. Buahnya diketahui<br />mengandung minyak atsiri yang didominasi senyawa trans-anethol (90%)<br />dan berkhasiat sebagai antiseptik, antispasmodik, antikanker, karminatif,<br />pelega tenggorokan, obat bronkitis, serta digunakan dalam pembuatan<br />sabun, parfum, pasta gigi, juga krim kulit. Sebagai tanaman bernilai<br />ekonomi, upaya perbanyakan anis perlu dilakukan. Perbanyakan secara in<br />vitro dengan teknik kultur jaringan merupakan salah satu metode alternatif<br />yang dapat digunakan untuk menghasilkan bibit dalam jumlah banyak,<br />seragam dan dalam waktu yang relatif singkat. Dengan penambahan<br />sitokinin sintetik tipe urea seperti thidiazuron (TDZ) dan tipe purin seperti<br />benzil amino purin (BAP) akan memacu inisiasi dan proliferasi tunas.<br />Penelitian ini bertujuan mendapatkan media yang tepat untuk menginduksi<br />tunas anis yang optimal dengan penambahan BAP atau TDZ, mengetahui<br />respon pertumbuhan dan penampakan kultur akibat penambahan berbagai<br />konsentrasi BAP atau TDZ, serta mempelajari sinergisme yang terjadi<br />antara keduanya. Pada tahap inisiasi, eksplan berupa tunas pucuk diinduksi<br />di dalam media MS padat dengan penambahan BAP (0,1 mg/l; 0,2 mg/l;<br />0,3 mg/l; 1 mg/l; 2 mg/l; 3 mg/l), atau TDZ dengan kisaran konsentrasi<br />yang sama. Tunas terbanyak yang dihasilkan dari dua jenis sitokinin pada<br />tahap ini disubkultur ke dalam media yang ditambahkan jenis sitokinin<br />yang berbeda (TDZ ke BAP atau BAP ke TDZ) pada konsentrasi 0,3 mg/l<br />atau 3 mg/l. Pada media yang ditambahkan TDZ dihasilkan tunas anis<br />lebih banyak (3,62-6,28) dibandingkan pada media yang ditambahkan<br />BAP (1,86-2,78), tetapi tunas yang dihasilkan pendek (roset). Sedangkan<br />tunas yang dihasilkan dalam media yang ditambahkan BAP beruas lebih<br />tinggi tetapi jumlah tunasnya sedikit. Subkultur tunas anis ke dalam media<br />yang diperkaya dengan sitokinin yang berbeda meningkatkan jumlah tunas<br />yang berproliferasi dan memperbaiki visual tunas.<br />Kata kunci: Anis, Pimpinellla anisum L. ,  minyak atsiri, multiplikasi tunas,<br />in vitro, TDZ, BAP, Jawa Barat<br />ABSTRACT<br />Application of purine and urea types of cytokinins in<br />shoot multiplication of Anise (Pimpinella anisum L.) in<br />vitro<br />Pimpinella anisum L. or sweet anise is an annual–herbaceous plant<br />belongs to the Umbelliferae family. The fruit of anise contains of essential<br />oil, which is mainly consisted of trans-anethol (90%). Essential oils of<br />anise is mainly used as an antiseptic, antispasmodic, anticancer,<br />carminative, expectorant and has also been used as component in soap,<br />perfumery, tooth paste, and skin cream productions. Since this crop is<br />mainly cultivated in sub tropical region, anise cultivation in Indonesia has<br />not been performed. To obtain sufficient numbers of anise planting<br />materials in vitro propagation was conducted by applying benzyl amino<br />purine (BAP) and thidiazuron (TDZ). In this research TDZ or BAP were<br />applied at various concentrations (0,1 mg/l: 0.2 mg/l; 0.3 mg/l; 1 mg/l; 2<br />mg/l; 3 mg/l), to induce shoots in MS-solid culture media. The highest<br />number of shoots obtained in those two type of cytokinins containing<br />media from the initiation stage were subcultured into the media<br />supplemented with different cytokinins (TDZ to BAP or BAP to TDZ) at<br />0.3 mg/l or 3 mg/l levels. The results showed that medium with the<br />addition of TDZ resulted in higher numbers of shoot (3,26-6,28) than that<br />of medium with an addition of BAP (1,86-2,78). However, rosette shoots<br />were dominant in TDZ containing medium. On the other hand, medium<br />with an addition of BAP resulted in less numbers of shoots with taller<br />nodes. Subculture of anise into different kinds of cytokinins increased the<br />numbers of proliferated-shoots and recovered the abnormal shoots.<br />Key words : Anise, Pimpinellla anisum L, essential oils, shoots<br />multiplication, in vitro, TDZ, BAP, West Java

scholarly journals Characterization of associative diazotrophic bacteria in torch ginger

2020 ◽  
Vol 41 (6) ◽  
pp. 2815-2824
Author(s):  
Alexandre José de Oliveira ◽  
◽  
Thaís Cristina Franco ◽  
Ligiane Aparecida FLorentino ◽  
Paulo Roberto Corrêa Landgraf ◽  
...  
Keyword(s):  
Bacterial Growth ◽  
Culture Media ◽  
Bacterial Isolates ◽  
Diazotrophic Bacteria ◽  
Bacterial Strains ◽  
Rock Powder ◽  
The Media ◽  
Semi Solid ◽  
Indole 3 Acetic Acid

Associative diazotrophic bacteria perform several processes that promote increased plant development and production, allowing a reduction in the use of agricultural inputs and costs. However, for some species, such as torch ginger, there are still no reports of studies aimed at identifying diazotrophic bacteria associated with this species. On this basis, this study proposes to isolate and characterize associative diazotrophic bacteria in rhizospheric soils and roots of torch ginger as well as analyze the potential of these isolates in solubilizing phosphorus (P) and potassium (K) and producing indole-3-acetic acid (IAA). Soil and roots samples of torch ginger were inoculated into five different semi-solid and semi-selective culture media, namely, NFb, JNFb, LGI, JMV and FAM, where bacterial growth was diagnosed by the formation of a characteristic film on the surface of the media. Subsequently, the bacterial isolates were analyzed for their ability to solubilize P and K in liquid medium, using phosphate rock powder (AO-15) and potassium rock powder (phonolite) as sources of P and K, respectively. All culture media showed bacterial growth, making this the first report of isolation of diazotrophic bacterial strains in this species. Eight of the obtained strains originated from rhizospheric soils and four from roots of torch ginger. Of these, 10 solubilized P, with the UNIFENAS 100-340, UNIFENAS 100-342 and UNIFENAS 100-348 strains standing out. Six strains showed K solubilizing ability, UNIFENAS 100-346 being the most efficient. All strains were able to produce the IAA phytohormone, both in the presence and absence of tryptophan, with superior results obtained by UNIFENAS 100-344 and UNIFENAS 100-351.

scholarly journals Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable Mycobacterium tuberculosis

2021 ◽  
Vol 10 (15) ◽  
pp. 3249
Author(s):  
Annelies W. Mesman ◽  
Seung-Hun Baek ◽  
Chuan-Chin Huang ◽  
Young-Mi Kim ◽  
Sang-Nae Cho ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Culture Media ◽  
Multidrug Resistant ◽  
Culture Conditions ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Solid Culture ◽  
Lowenstein Jensen ◽  
Genetic Mechanisms ◽  
Lipid Resuscitation

An estimated 15–20% of patients who are treated for pulmonary tuberculosis (TB) are culture-negative at the time of diagnosis. Recent work has focused on the existence of differentially detectable Mycobacterium tuberculosis (Mtb) bacilli that do not grow under routine solid culture conditions without the addition of supplementary stimuli. We identified a cohort of TB patients in Lima, Peru, in whom acid-fast bacilli could be detected by sputum smear microscopy, but from whom Mtb could not be grown in standard solid culture media. When we attempted to re-grow Mtb from the frozen sputum samples of these patients, we found that 10 out of 15 could be grown in a glycerol-poor/lipid-rich medium. These fell into the following two groups: a subset that could be regrown in glycerol after “lipid-resuscitation”, and a group that displayed a heritable glycerol-sensitive phenotype that were unable to grow in the presence of this carbon source. Notably, all of the glycerol-sensitive strains were found to be multidrug resistant. Although whole-genome sequencing of the lipid-resuscitated strains identified 20 unique mutations compared to closely related strains, no single genetic lesion could be associated with this phenotype. In summary, we found that lipid-based media effectively fostered the growth of Mtb from a series of sputum smear-positive samples that were not culturable in glycerol-based Lowenstein–Jensen or 7H9 media, which is consistent with Mtb’s known preference for non-glycolytic sources during infection. Analysis of the recovered strains demonstrated that both genetic and non-genetic mechanisms contribute to the observed differential capturability, and suggested that this phenotype may be associated with drug resistance.

Integration of culture-dependent and independent methods provides a more coherent picture of the pig gut microbiome

2020 ◽  
Vol 96 (3) ◽  
Author(s):  
Gavin J Fenske ◽  
Sudeep Ghimire ◽  
Linto Antony ◽  
Jane Christopher-Hennings ◽  
Joy Scaria
Keyword(s):  
Bacterial Communities ◽  
Bacterial Species ◽  
Shotgun Metagenomics ◽  
Culture Techniques ◽  
Microbiome Composition ◽  
Culture Independent ◽  
Health And Disease ◽  
The Media ◽  
And Function ◽  
Culture Dependent

ABSTRACT Bacterial communities resident in the hindgut of pigs, have profound impacts on health and disease. Investigations into the pig microbiome have utilized either culture-dependent, or far more commonly, culture-independent techniques using next generation sequencing. We contend that a combination of both approaches generates a more coherent view of microbiome composition. In this study, we surveyed the microbiome of Tamworth breed and feral pigs through the integration high throughput culturing and shotgun metagenomics. A single culture medium was used for culturing. Selective screens were added to the media to increase culture diversity. In total, 46 distinct bacterial species were isolated from the Tamworth and feral samples. Selective screens successfully shifted the diversity of bacteria on agar plates. Tamworth pigs are highly dominated by Bacteroidetes primarily composed of the genus Prevotella whereas feral samples were more diverse with almost equal proportions of Firmicutes and Bacteroidetes. The combination of metagenomics and culture techniques facilitated a greater retrieval of annotated genes than either method alone. The single medium based pig microbiota library we report is a resource to better understand pig gut microbial ecology and function. It allows for assemblage of defined bacterial communities for studies in bioreactors or germfree animal models.

scholarly journals The Roles of Porous Coral Sands in Initial Enrichment of Ammonia-Oxidizing Bacteria

2007 ◽  
Vol 7 ◽  
pp. 525-532
Author(s):  
Qing Guo ◽  
Zao-he Wu ◽  
Ming-liang Qian ◽  
Binhe Gu
Keyword(s):  
Culture Media ◽  
Potassium Dihydrogen Phosphate ◽  
N:P Ratio ◽  
Ammonium Nitrogen ◽  
Dihydrogen Phosphate ◽  
Ammonia Oxidizing Bacteria ◽  
Potassium Dihydrogen ◽  
Conversion Rates ◽  
Calcium Source ◽  
The Media

The purpose of this study was to investigate the roles of coral sands in the enrichment and isolation of ammonium-oxidizing bacteria (AOB). We hypothesized that the porous coral sands provided additional surface area and nutrients for the growth of periphytic AOB. In the present study, an orthogonal test was designed to compare the AOB conversion rates of ammonium-nitrogen (NH4+N) to nitrite-nitrogen (NO2--N) among various combinations of culture media. Results showed that the conversion of NH4+N to NO2--N increased significantly when the coral sands were added, implying that coral sands were beneficial to the growth of AOB. Additions of potassium dihydrogen phosphate (KH2PO4) or sodium bicarbonate (NaHCO3) to the media became unnecessary when coral sands were used, but the addition of KH2PO4was needed when the molar nitrogen to phosphorus (N:P) ratio reached 10 in the enrichment media using calcium carbonate (CaCO3) powder as a calcium source.

In vitro evaluation of the effect of nicotine, cotinine, and caffeine on oral microorganisms

2008 ◽  
Vol 54 (6) ◽  
pp. 501-508 ◽  
Author(s):  
Karina Cogo ◽  
Michelle Franz Montan ◽  
Cristiane de Cássia Bergamaschi ◽  
Eduardo D. Andrade ◽  
Pedro Luiz Rosalen ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Culture Media ◽  
Bacterial Species ◽  
In Vitro Study ◽  
Bacterial Strains ◽  
Planktonic Cells ◽  
Susceptibility Tests

The aim of this in vitro study was to evaluate the effects of nicotine, cotinine, and caffeine on the viability of some oral bacterial species. It also evaluated the ability of these bacteria to metabolize those substances. Single-species biofilms of Streptococcus gordonii , Porphyromonas gingivalis , or Fusobacterium nucleatum and dual-species biofilms of S. gordonii – F. nucleatum and F. nucleatum – P. gingivalis were grown on hydroxyapatite discs. Seven species were studied as planktonic cells, including Streptococcus oralis , Streptococcus mitis , Propionibacterium acnes , Actinomyces naeslundii , and the species mentioned above. The viability of planktonic cells and biofilms was analyzed by susceptibility tests and time-kill assays, respectively, against different concentrations of nicotine, cotinine, and caffeine. High-performance liquid chromatography was performed to quantify nicotine, cotinine, and caffeine concentrations in the culture media after the assays. Susceptibility tests and viability assays showed that nicotine, cotinine, and caffeine cannot reduce or stimulate bacterial growth. High-performance liquid chromatography results showed that nicotine, cotinine, and caffeine concentrations were not altered after bacteria exposure. These findings indicate that nicotine, cotinine, and caffeine, in the concentrations used, cannot affect significantly the growth of these oral bacterial strains. Moreover, these species do not seem to metabolize these substances.

scholarly journals Neutral-Red in the Routine Examination of Water

1902 ◽  
Vol 2 (3) ◽  
pp. 314-319 ◽  
Author(s):  
Ernest E. Irons
Keyword(s):  
Organic Dyes ◽  
Culture Media ◽  
Bacterial Species ◽  
Neutral Red ◽  
Green Fluorescence ◽  
Routine Examination ◽  
Water Supplies ◽  
River Waters

The use of neutral-red culture media has recently been suggested for the detection of B. coli in water supplies. Rothberger (1) grew a number of organisms on media containing various organic dyes, and found that while some species were able to bring about a reduction of the colouring matter in the medium, others produced no change. This difference in reaction was particularly marked in the case of B. typhosus and B. coli in media containing neutral-red. B. coli casused the reduction of the neutral-red to yellow with green fluorescence, whilst B. typhosus produced no change of colour beyond an occasional fading of the red. After testing a number of races of B. coli and B. typhosus, Rothberger proposed neutral-red media for the differentiation of the two organisms. Scheffler (2) confirmed in the main the work of Rothberger, and further observed that the reaction is by no means specific for B. coli. In addition to a number of bacterial species not commonly found in water, which gave the reaction, Scheffler found 3 of 13 organisms in spring and river waters, and 8 of 18 intestinal organisms from man which, though not belonging to the colon group, gave the neutral-red reaction.

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