An improved method of dyeing of bakery yeast according to Gram

2020 ◽  
pp. 44-50
Author(s):  
P. Kulyasov ◽  
A. Bekieva ◽  
B. Nayminov ◽  
B. Gilgeev ◽  
B. Zodbaev
Keyword(s):  
Biological Sciences ◽  
Methylene Blue ◽  
Gentian Violet ◽  
The Body ◽  
Yeast Cultures ◽  
Direct Acquaintance ◽  
Lugol Solution ◽  
Modern Level ◽  
Bakery Yeast ◽  
Improved Technique

Knowledge and practical skills of working with microorganisms are required for mastering the modern level of development of veterinary and biological sciences. Microorganisms are the main objects of biotechnology, molecular biology and genetics, and they constantly accompany human and animals in the environment and cohabit inside the body. Direct acquaintance with them and the development of the principles of microbiological research will not only improve their professional level, but also to acquire knowledge and skills are not superfl uous in everyday life. The microscopic method involves the study of living or killed representatives of the microbial (bacteria, bacilli, clostridiums and plectridia) and yeast (bakery, fruit and vegetable, berry and grape yeast) world in a colored or unpainted state using a binocular light microscope. An improved technique of the famous Danish microbiologist G. K. Gram for coloring bakery’s yeast has been presented in the article. On the basis of laboratory researches it has been shown that the diff erentiated coloring according to Gram can be changed on color scale, depending on dyes. Classically according to Gram used: gentian violet, Lugol solution, fuchsin Pfeiff er (based on carbolic fuchsin), but in our technique instead of gentian violet methylene blue has been used, Lugol solution, instead of fuchsin Pfeiff er took red safranin and additionally in the manufacture of the third drug – diamond green dye. It has been shown that the coloring of microbial or yeast cultures has being achieved through the latter dye. So, for example with methylene blue, microbes or yeast are colored blue, with safranin in red and with diamond green in green.

scholarly journals THE EFFECT OF GENTIAN VIOLET ON PROTOZOA AND ON TISSUES GROWING IN VITRO, WITH ESPECIAL REFERENCE TO THE NUCLEUS

1914 ◽  
Vol 20 (6) ◽  
pp. 545-553 ◽  
Author(s):  
D. G. Russell
Keyword(s):  
Methylene Blue ◽  
Bacterial Contamination ◽  
Gentian Violet ◽  
Bactericidal Effect ◽  
Adult Tissue ◽  
Animal Cells ◽  
Selective Action ◽  
Especial Reference ◽  
Made In

1. Gentian violet may be regarded as a true vital nuclear stain. 2. Embryonic and adult tissue of the frog will grow in vitro in the presence of gentian violet of a far stronger dilution than that necessary to kill many bacteria. In these experiments, for example, successful tissue growths were obtained when gentian violet 1 to 20,000 was used, yet Bacillus subtilis will not grow in 1 to 100,000 dilution and grows very badly in 1 to 1,000,000 dilution. This fact may simplify the technique of the growth of certain tissues by eliminating the risk of bacterial contamination. 3. The use of stains in the plasma in which tissue is grown will probably facilitate the study of nuclear growth. 4. Gentian violet appears to have a certain selective action on tissue similar to that exercised by the dye on bacteria. Certain observations made last year in this laboratory (too few to serve as more than a suggestion) seem to indicate that another dye (methylene blue) acted as a stimulant to the growth of connective tissue. These leads should be followed out and the effect of various stains studied in the hope of discovering dyes which will exercise a sharp selective action on growing tissue. 5. The growth of animal cells in a strength of dye much more than sufficient to kill many pathogenic organisms lends encouragement to efforts now being made in this laboratory to apply the observations on the bactericidal effect of gentian violet and allied stains to the treatment of disease. Moreover, the ability of growing cells to rid themselves of this dye would indicate that it may be possible to apply the dye to infected tissue and count on the cells to eliminate the material after its work had been done.

214. Studies in mastitis. VI. General observations, summary and conclusions

1939 ◽  
Vol 10 (1) ◽  
pp. 104-107 ◽  
Author(s):  
J. G. Davis ◽  
J. McClemont
Keyword(s):  
Methylene Blue ◽  
Chemical Composition ◽  
Selective Medium ◽  
False Positives ◽  
The Body ◽  
General Idea ◽  
Blood Agar ◽  
Total Count ◽  
Confirmatory Test ◽  
Paper Strip

I. Methods of taking and examining samples of milk for Str. agalactiae are described.Where delay in examination is unavoidable Edwards's (1) medium is recommended. This selective medium, however, does not permit the growth of some types of organisms which may cause a subclinical mastitis. It is therefore recommended that, where possible, samples be examined immediately after taking and a non-selective medium used. In this way a measure of the udder count is obtained which is of great value. Where samples can be examined within a few hours plain blood agar is preferred. If blood is unobtainable ordinary milk agar may be used. Non-blood media have certain advantages over blood media, which compensate for the loss of ability to detect haemolysis when it occurs.A simple confirmatory test for Str. agalactiae is described. Confirmation of type is regarded as essential in all but the obvious cases.II. The results of a number of simple tests for mastitis have been compared with those from examination in blood agar. These all have roughly the same order of efficiency in that they detect about half the positive cases and give 10–30 % false positives. It is suggested, therefore, that with two exceptions (the milk agar count and the rennet test, which may be used for special purposes) they are so unreliable as not to be worth doing in any laboratory examination.The brom-cresol-purple paper, strip cup and induration tests, which are very simple and can be carried out by the milker, are worth doing in order to get a general idea of the incidence of mastitis in the herd. Their limitations should be realized, however, for no simple test, or combination of simple tests, is really satisfactory, especially if the owner wishes to secure eradication of the disease from the herd. Our results thus confirm those of other workers. III. Mastitis has a marked effect on the “udder count” as given by total colony count on milk agar. No samples from mastitis-free cows were found to have a count of over 2000 per ml. and very few over 500. About 50% of infected cows yielded milk with a count of over 1000.The milk agar count is perhaps the best of the indirect tests as although it detects no more than other tests (about 50 %) it gives very few false positives. A count of under 100 per ml. may be taken to indicate freedom from infection and a count of over 1000 to indicate mastitis.IV. The methylene blue reduction test is not capable of detecting mastitis. Reduction time appears to be more closely related to the cell content of infected milks than to the total count. Storage of the samples for 16 hr., either at 4 or 15·5° C, did not materially improve the efficiency of the test.V. The possible causes of the unsuitability of mastitis milk for cheesemaking are classified and discussed. The brom-cresol-purple-rennet test is described and recommended as a general test for mastitis and suitability of the milk for cheese-making. The effect of mastitis on the chemical composition and enzyme content of the milk is discussed. Mastitis adversely affects the “body” of cheese and is an important factor in the fault known as “red spot”. Many samples of mastitis milk give slow growth of starter, and these are usually detected by the rennet test. Deficiency in bacterial growth factors may be one factor in causing slow starter. This phenomenon is also correlated with rapid reduction of methylene blue and high total count, but the real factor responsible is probably abnormal chemical composition.VI. The incidence of mastitis increases steadily with age of the animal. Older cows do not give milk of higher “udder count” than younger cows if infection is absent. The increased count is entirely due to the increased incidence of mastitis. There appears to be a somewhat lower udder count in late lactation in infected cows, but a slightly higher one if mastitis is absent.Finally we may emphasize the fact that for eradication purposes there are only two methods of diagnosis worth using—simple tests on the farm and carefully controlled bacteriological examination in the laboratory.

scholarly journals RESISTANCE OF BACTERIAL SPORES TO THE TRIPHENYLMETHANE DYES

1925 ◽  
Vol 41 (4) ◽  
pp. 471-477 ◽  
Author(s):  
Victor Burke ◽  
Horace Skinner
Keyword(s):  
Bacillus Anthracis ◽  
Pathogenic Bacteria ◽  
Gentian Violet ◽  
The Body ◽  
Bacterial Spores ◽  
Bactericidal Action ◽  
Triphenylmethane Dyes ◽  
Vegetative Cells ◽  
Bacteriostatic Action ◽  
Original Culture

1. Bacterial spores are highly resistant to the bactericidal action of the triphenylmethane dyes. Many Bacillus anthracis and Bacillus subtilis spores resist a saturated aqueous solution of gentian violet for 24 hours at 37°C. They also resist exposure to the same dye solution for 10 minutes at 80°C. 2. The selective bactericidal action of these dyes applies only to the vegetative cells. Spores of the Gram-positive bacteria are more resistant to these dyes than the vegetative cells of Gram-negative bacteria. 3. The vegetative cells of Bacillus anthracis show variation in resistance to the bacteriostatic action of gentian violet. The spores of Bacillus anthracis show variation in resistance to the bactericidal action of gentian violet. 4. Bacillus anthracis can increase in resistance to the bacteriostatic action of gentian violet and grow in dye dilutions inhibiting the original culture. There is the possibility of pathogenic organisms becoming dye-resistant in the body when exposed to non-bactericidal concentrations. For this reason it is advisable to use the greatest concentration of dye compatible with tissue tolerance. 5. Since the spores of pathogenic bacteria may lie dormant for longer periods than the dyes retain their bacteriostatic action in the body frequent applications of the dye should be made in preventing infections by spore-bearing bacteria.

THE INFLUENCE OF THE PHYSICAL ENDURANCE OF A PERSON

2020 ◽  
Author(s):  
Yana Vladimirovna Kizilova ◽  
Keyword(s):  
Biological Sciences ◽  
Living Conditions ◽  
The Body ◽  
Physical Culture ◽  
Physical Endurance ◽  
Internal Organs

The basis of physical culture - methodical and biological Sciences. Man differs from the rest of the fauna developed thinking, speech, the structure of internal organs and the specifics of living conditions. It is impossible to make a technique for training a person without knowing all aspects of the functioning of the body systems.

CORD-LIKE BACTERIA – A LIVING BIOLOGICAL CAUSATIVE AGENT OF MALIGNANT CANCER

2020 ◽  
Vol 15 (6) ◽  
pp. 841-855
Author(s):  
P.A. Kulyasov ◽  
Keyword(s):  
Mammary Gland ◽  
Gentian Violet ◽  
Nerve Impulse ◽  
Optical Microscope ◽  
The Body ◽  
Blue Dye ◽  
Improved Method ◽  
Living Body ◽  
Von Liebig ◽  
Pathological Material

In the study of the malignant cancerous thickening of the tumor-like nature in a cat’s mammary gland with the subsequent staining of the pathological material by the improved method according to H. C. Gram, a category of rod-shaped bacteria of the cord form was found. It was noted that these microscopic (visible in an optical microscope) bacteria are able to cluster into dense formations and, with an increase in the temperature of the external environment, form dense living “ropes” and “bundles” from tens of thousands microbial individuals. This gives them the opportunity, getting and reproducing in a favorable environment, thereby increasing their population in quantity, inside the living body of the diseased animal organism, pulling the affected parts of the body and its internal organs and tissues with cord-like overlays, leading to their microbial suffocation. The flow of nutrient and protective elements with the blood stops and the transmission of the nerve impulse is disrupted, which immediately leads the affected parts of the body to tumor-like growths with their subsequent necrosis (necrosis). Dead tissue, and in our case, the mammary gland of a cat, is an excellent breeding ground for the development and reproduction of colonies of ciliated microbes in it, capable of forming spores in case of unfavorable factors for their vital activity. Reproducing and becoming sexually mature individuals, within 20 hours (approximately, in time, less than one day), cord-like microbes begin to immediately form living microbial bundles of various thicknesses and sizes. Covering a certain separate area of living tissue with a microbial “loop”, squeezing the affected parts of a living body – blood vessels and nerves, thereby slow down, and in some cases completely stop blood flow and nerve impulse. All these negative factors gradually lead to the extinction of the vital tone of muscle and nervous tissues, which inevitably leads to cell death and the growth of a dense tumor-like neoplasm. Malignant cancerous tumor of domestic animals does not arise from the previously voiced chemical theory of Justus von Liebig, but because of the presence of a living biological pathogen – ciliate rod-shaped formed bacteria – inside the living parts of the body. Staining according to the improved method of H. C. Gram with blue dye (methylene blue) instead of a violet dye gentian violet, and a red dye (safranin) instead of Pfeiffer's carbolic fuchsin, allowed under the optical immersion system of a light binocular microscope with a magnification of 1000 times, to see special cord-like, wriggling loop-shaped forms, consisting of a large number of rod-shaped microbes. They are able to create inseparable twisted chains and “loops” from their tiny microbial bodies, which gives grounds to assert their significant role in the occurrence of malignant cancerous neoplasms.

scholarly journals The hydrogen-ion concentration and the oxidation-reduction potential of the cell-interior: a micro-injection study

1925 ◽  
Vol 98 (689) ◽  
pp. 259-286 ◽  
Keyword(s):  
Methylene Blue ◽  
Gentian Violet ◽  
Reducing Power ◽  
Ion Concentration ◽  
Hydrogen Ion Concentration ◽  
Oxidation Reduction ◽  
Oxidation Reduction Potential ◽  
Living Organisms ◽  
Actual Formation ◽  
Qualitative Estimation

It has been recognised since the middle of the eighteenth century that one of the most fundamental characteristics of living organisms is their capacity to oxidise substances incapable of oxidation at ordinary temperatures; but no qualitative estimation of this power of oxidation was carried out until the time of Ehrlich, whose classical experiments on the injection of methylene blue into the intact animal revealed the fact that certain organs seemed to have a higher reducing power than others. Later, much histo-chemical work was done. Numerous observers studied the effect of staining tissues and cells in reagents which indicated by their colour whether they were reduced or oxidised. Several such indicators were used by the earlier workers, especially alphanaphthol and pyronin and alpha-naphthol and gentian violet; but the two chief methods were the intracellular formation of an indophenol, introduced by Röhmann and Spitzer in 1895 (20), and the oxidation of the leucobase of methylene blue, introduced by Unna in 1911 (23). In the latter case the cell was placed in a solution of the completely reduced dye, and the conclusion was that wherever the blue colour appeared, there the cell had been able to oxidise it. The indophenol method depended on the actual formation and precipitation of a dye in the cell by an oxidative condensation. The original reactants used were dimethylparaphenylenediamine and alpha-naphthol, but various later workers modified this by using other phenols and other aromatic amines, so that indophenols of different colours were produced.

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