Casein Digest Medium Research Articles

Suitability in compounding sterile preparations: An observational study in a referral hospital

Context: Compounding sterile preparations in hospitals requires special attention because the process is more complex. Errors that occur when compounding sterile preparations can cause contamination of the prepared preparations. Aims: To evaluate the compounding of sterile preparations in a referral hospital. Methods: This research was conducted by observing nine critical aspects in the compounding of sterile preparations. The research instrument used was a checklist, as regulated in USP <797> Guidelines and Basic Guidelines for Dispensing Sterile Preparations. After that, the most mixed preparations were tested for sterility using a fluid thioglycollate medium and soybean casein digest medium. Results: Of the 36 compounding sterile preparations, discrepancies were found in the aspects of compounding personnel (100%), building (100%), equipment (100%), aseptic procedures (100%), packaging (0%), labels (100%), storage (2.78%), distribution (2.78%) and quality assurance (100%). Furthermore, sterile preparations prepared in the cleanroom in this referral hospital indicate the occurrence of microbial contamination. Conclusions: Most of the critical aspects in compounding sterile preparations at this referral hospital have not met the recommendations of the USP <797> Guidelines and the Basic Guidelines for the Dispensing of Sterile Preparations, except for the packaging aspect. Improvements in several critical aspects need to be made to ensure the quality of the prepared preparations from microbial contamination.

Investigation of low-cost media for Bacillus thuringiensis subspecies israelensis

Bacillus thuringiensis subspecies israelensis (Bti) has been used as a mosquito control agent in agricultural field and residences. The production of Bti by conventional medium was expensive. The objective of this study was to investigate low-cost media for Bti’s production with a cost analysis of the medium formulated. Soybean Casein Digest Medium (TSB) was used as a conventional medium to compare with Saberi medium at room temperature (approximately 21.5˚C) on static and agitation (by obituary shaker at 100 rpm). Saberi medium was also modified to vary the percentage of nitrogen and carbon source to observe the growth rate and sporulation, compared to the Saberi medium. The growth of Bti in TSB with agitation for 24 hours yielded the highest growth rate. Sporulation of Bti in TSB with agitation at room temperature was 6 days which was faster than Bti in the Saberi medium. Modified Saberi nitrogen source and carbon source medium could not support the growth of Bti due to an imbalance in the percentage of nitrogen source and carbon source. Determination of sugar utilization showed sugar in the medium was utilized mostly in the exponential phase of Bti, from 3 hours to 6 hours. The price of TSB was 76.80 Thai baht per liter while Saberi medium’s cost was 20.43 Thai baht per liter. In this study, the Saberi medium may not be able to replace TSB in terms of growth rate and period of sporulation. Further study will be needed to optimize the media that use agro-industrial wastes as ingredients following the concept of circular economy and sustainability.

Antibiotic Activity of Actinomycetes Isolated from Young Tectona Grandis (L.) Wood and Pith

Actinomycetes are a source for novel bioactive compounds and justify obtaining new species from various sources. Hardwoods such as Tectona grandis (L.) have not been studied for actinomycete isolation. We aim to isolate endophytic actinomycetes from young Tectona grandis wood and pith and screen for antibiotic activity. Five young wood were cut and surface sterilized using ethanol and hypochlorous acid. The wood and pith of each sample are placed in eight plates of Humic acid-vitamin B (HV), Tap water Yeast extract (TWYE), and Yeast Extract Casein Digest (YECD) medium and incubated at 27°C for four weeks. Actinomycetes were isolated from such medium, observed every week, and transferred to an International Streptomyces Project-2 (ISP-2) medium for identification and antibiotic production tested against Staphylococcus aureus, Helicobacter pylori, and Escherichia coli using the Kirby-Bauer method. Seven actinomycetes were isolated from the wood, primarily from YECD and TWYE media, with varying morphological characteristics. One isolate having maroon-colored aerial and vegetative mycelium with grey spores showed moderate antibacterial activity against S. aureus and H. pylori (13.49±1.03 mm and 14.9±0.7 mm, respectively), while two other actinomycetes showed weak activity against these bacteria. However, none of the actinomycetes show any activity against E. coli. Tectona grandis (L.) is a potential source for novel actinomycetes with an antibiotic activity which warrants further exploration

Microbial Growth in Soyabean Casein Digest Medium and its Interpretation

Microbial Growth in Soyabean Casein Digest Medium and its Interpretation

Microbiological studies of a new combined drug “Proplantmed” based on propolis phenolic hydrophobic drug

Gastric and duodenal ulcer is one of the most widespread diseases in the world; therefore, creation and research of new effective and safe medicines are of great importance.Aim. To study the microbiological properties of the new combined drug “Proplantmed” based on propolis phenolic hydrophobic drug (PPHD).Materials and methods. The studies of microbiological purity of the drug were performed according to the State Pharmacopoeia of Ukraine (SPhU). The following growth media were used for testing: soybean casein digest medium (“HIMedia Laboratories Pvt. Ltd. India”), soybean casein digest agar (“HIMedia Laboratories Pvt. Ltd. India”), thioglycollate medium for sterility control (“HIMedia Laboratories Pvt. Ltd. India”) and Sabouraud dextrose agar (manufacturing country: India, (“HIMedia Laboratories Pvt. Ltd. India”)).Results. The experimental samples of the drug “Proplantmed” meet the requirements of the SPhU and “Microbiological Purity” test; this fact is an important quality indicator of the finished medicinal product.Conclusions. According to the results of the experiment conducted it has been found that level of microbiological purity of the granules proposed meets the requirements of the SPhU; this indicator is the quality guarantee of this drug.

OPTIMIZATION OF PHYSICAL PARAMETERS OF ?-AMYLASE PRODUCING BREVIBACILLUS BOROSTELENSIS R1 IN SUBMERGED FERMENTATION

Bacteria have been regarded as treasure of many use ful enzymes viz., amylases, proteases, lipases, hyd rolases and reductases. Among them amylolytic enzymes have great biotechnological applications and economic exploitations. The produ ction of α-amylases by fermentation had been thoroughly investigated and s hown to be affected by a variety of physicochemical factors, such as the composition of the growth medium, the type of strai n, cell growth, methods of cultivation, inoculum co ncentration, time of incubation, pH, temperature, salinity, carbon, nitrogen and min eral sources. The present study was carried out to optimize the α-amylase production of Brevibacillus borstelensis R1 using t en different media viz., Nutrient broth, Luria Bert ain broth, Clarks & Lub medium, Pikovskaya’s medium, Tendler's non-synthetic medium , Amylase production medium, Soluble starch beef ex tract medium, Soybean casein digest medium, Yeast extract peptone dextros e glucose medium and Tryptone glucose beef extract medium. Among these ten media, Pikovskaya’s (PK) medium proved to be optima l for α-amylase production (1861±17U/ml). The optimized α-amylase production in PK medium by submerged fermentation ( SmF) was subjected to varying physical parameters s uch as 24hrs incubation time, 2% inoculum size, 37 0 C, pH 7.0 and 1% NaCl. Alpha-amylase produced by B. borostelensis R1 have many applications in starch processing, desizing of textiles, paper sizing, detergent additive, bread improvement, ethanol product ion, sewage treatment, effluent treatment and other fermentation processes.

Alkaline protease from Bacillus cereus VITSN04: Potential application as a dehairing agent

The objective of this work is to use protease enzyme as an ecofriendly alternative to chemicals in dehairing. An alkaline protease producing bacterium was isolated from protein-rich soil sample. The bacterium was identified as Bacillus cereus VITSN04 by 16S rRNA gene sequencing method. Growth characteristics and protease activity were studied in yeast, malt, beef, nutrient broth and soybean casein digest media and the enzyme secretion was found to correspond with growth. Maximum protease production was obtained in soybean casein digest medium at 16h with the activity of 200.1±0.68U/ml and a correlation coefficient of 0.965 between growth and enzyme production. The crude enzyme was found to have maximum activity at 30°C and pH 8.0. The protease was purified by ammonium sulphate precipitation, Sephadex G-50 and G-100 gel filtration chromatography. The purified protease was homogeneous on non-denaturing PAGE and its molecular weight was estimated to be 32kDa. The purified protease was of the serine type as it was inhibited by phenylmethylsulphonyl fluoride. The crude enzyme preparation was found to be effective in dehairing goat skins in leather processing.

Isolation and morphological characterization of antibiotic producing actinomycetes

Purpose: To isolate and characterize antibiotic producing actinomycetes from soil samples in Belgaum, Karnataka, India. Methods: Crowded plate technique was used for the isolation of actinomycetes in media such assoybean – casein digest medium and actinomycetes isolation agar. The morphological and cultural characterization of one of the selected strains, designated A-4, was performed as per International Streptomycete Project (ISP). Results: Morphological and cultural studies showed that A-4 belonged to the Actinomycete genus. The morphological and cultural characteristics of the A-4 mutant showed cellular and aerial growth as well as soluble pigment formation in various ISP media. Conclusion: Findings from this investigation revealed that the selected strain, A-4, is an actinomycete.Keywords: Actinomycete, Antibiotic, MIC, Crowded plate technique, Morphology

Studies on rheology and microbiological evaluation of hydrotropically gelled starch as topical vehicle for terbinafine hydrochloride

The present study was aimed for the development and evaluation of hydrotropic starch gels of terbinafine hydrochloride which is an allylamine derivative of antifungal agent was formulated by using corn starch and sodium salicylate as hydrotropic salt. The gels were prepared in presence and absence of propylene glycol. The prepared gels were evaluated for in vitro drug release, rheological behavior and microbial studies. The degree of increase in the drug diffusion was found to be in order corn starch gel with propylene glycol was greater than corn starch gel without propylene glycol. The microbial studies were carried out in soya bean casein digest medium with Candida albicans as test organisms and were found to be 22.86 ± 0.58 and 20.22 ± 0.65 mm for TCSG (IV) and TCS (III), respectively. All the gels exhibited shear thinning. The rheogram indicated that the gel systems are pseudoplastic and exhibited thixotropy. The added propylene glycol has not appreciably altered the apparent viscosity values.

Partial replacement of succinate dehydrogenase function by phage- and plasmid-specified fumarate reductase in Escherichia coli.

Phages capable of transducing succinate dehydrogenase mutants (sdh) of Escherichia coli were isolated from pools of artificially constructed recombinant lambda phages using a selective casein digest medium. These phages produced characteristically dense turbid plaques, and as prophages they increased the aerobic growth efficiencies of sdh mutants on complex media but were unable to promote growth with succinate as sole carbon and energy source (an essential feature of sdh + strains). The phages were identified as fumarate reductase transducing phages (lambda frdA) by the presence of a characteristic 4.9 kilobase pairs R.HindIII fragment of bacterial DNA, the expression of a polypeptide with a relative molecular mass of 72,000 (the frdA gene product) and by comparing their transducing activities with authentic lambda frdA phages. In parallel studies a strain containing a ColE1-frd hybrid plasmid (pGS1 = pLC16.43) was characterized. Transfer of pGS1 to sdh mutants was accompanied by increased aerobic growth efficiencies on complex media and the ability to utilize succinate as sole carbon and energy source. It was concluded that fumarate reductase can replace succinate dehydrogenase but the extent of the reversal of the sdh lesion depends on frd gene dosage and the titration of the repressor which normally prevents aerobic synthesis of the reductase.

Increased spore yields of Clostridium perfringens in the presence of methylxanthines

The methylxanthines caffeine, theophylline, and isobutylmethylxanthine greatly increased spore yields of Clostridium perfringens strains FD-1, PS52, and PS49 when grown on Duncan-Strong medium or on a new casein-digest medium. Four other strains (KA3, and National Collection of Type Cultures strains 8798, 8238, and 10240) failed to show any significant increase when tested under similar conditions. The degree of sporulation increase was influenced by the carbohydrate energy source in some strains but not in others. Strain PS52 showed a large increase in spore yield when dextrin was the energy source but only a slight increase when raffinose served as the energy source. Strain FD-1 showed similar increases in spore yield with either dextrin or raffinose.

Comparative Study of Seven Media for Sterility Testing

A comparative study of the efficacy of sterility testing of pharmaceuticals according to two systems is described. The culture media used in the first system were: dithionite—thioglycollate (HS-T) both, recommended by the Nordic Pharmacopoeia Board, and a peptone liver digest medium or a peptone liver digest agar. The media used in the second system were: fluid thioglycollate medium and soybean—casein digest medium, both prescribed by the USP XVIII, and Sabouranud liquid medium or Sabouraud dextrose agar. The supplemental use of one of the two latter media has been recommended, because the USP XVIII media were considered inadequate. A total of 180 cultures of fastidious bacteria. Yeats, and molds was investigated. The first system was found superior to the second.

Growth of Escherichia coli NCTC 5928 in relation to enzymes in vitamin B6 metabolism.

RESULTS have been obtained which indicate the presence in Escherichia coli NCTC 86 of pyridoxamine phosphate oxidase1 and of pyridoxine phosphate oxidase2. These enzymes had previously been demonstrated in rabbit liver preparations3–6. These enzymes in E. coli NCTC 86 have been investigated in aqueous extracts obtained from acetone-ether dried cells (kindly supplied by the Lister Institute of Preventive Medicine, Elstree, Herts.), which had been grown for 18 h at 37° C on a casein-digest medium solidified with agar2.

413. The growth requirements of lactobacilli in relation to cheese flavour development

An attempt was made to devise a medium for the growth of lactobacilli which would simulate the nutritive conditions existing in ripening cheddar cheese. The basal medium finally used contained soluble digestion products of casein (formed by the action of rennet), lactose, lactate and phosphate. This medium gave a fair, but not a very heavy growth of flavour-producing lactobacilli isolated from cheddar cheese, and it was evidently deficient in growth-promoting power as compared with some media used in the laboratory. Additions of potassium and manganese salts to the casein digest medium gave a significant increase in growth. Potassium had the effect of off-setting the inhibition caused by the presence of an excess of sodium salts such as are normally present in cheese. The implications of the results in cheese ripening and their possible application in cheese manufacture are discussed.

A further investigation of the response of Leuconostoc to nicotinic acid and related compounds.

This report is an extension of an earlier study which dealt with certain unusual aspects of the growth response of Leuconostoc to nicotinic acid and related compounds. It was first reported by Johnson1 that Leuconostoc mesenteroides 9135, though requiring nicotinic acid, did not respond to ordinary amounts of nicotinamide or nicotinuric acid. It was found by the writers2 that of 6 Leuconostoc cultures which needed nicotinic acid for prompt growth in a casein digest medium, two of them failed to grow in the presence of nicotinamide, diphosphopyridine nucleotide (DPN), or triphosphopyridine nucleotide (TPN), while a third culture failed to grow in nicotinamide but grew promptly when DPN or TPN was supplied. Also, these 3 cultures did not respond to ordinary amounts of nicotinuric acid and methyl nicotinate, compounds which usually are utilized readily by other nicotinic acid requiring bacteria. The present work deals with certain aspects of the behavior of Leuconostoc not covered in our earlier report and includes also some tests with Streptococcus salivarius. The series of Leuconostoc cultures was extended to include additional representatives of the genus. A synthetic medium of amino acids and other definite compounds was used for this larger series of cultures in place of

Action of Tyrocidin and Detergents in Liberating Amino-Acids from Bacterial Cells

THE estimation of amino-acids by the amino-acid decarboxylase technique1 provides a means for the investigation of the amino-acid content of the internal environment of bacterial cells. If 1 ml. of a thick washed suspension of S. faecalis cells grown in casein digest medium is placed along with 1·5 ml. phosphate buffer pH 6·0 in the main cup of a Warburg manometer and 0·5 ml. lysine decarboxylase preparation tipped in from a side-bulb in the usual way, there is a small output of carbon dioxide of magnitude depending upon the degree of washing of the cells and corresponding to the amount of lysine carried down with and adsorbed on the surface of the cells (see curve labelled'intact cells'). If a further aliquot of cell suspension is treated in such a way as to disintegrate the cells, either by mechanical shaking with glass beads2 or by immersion in a boiling water bath for 10 min., before the addition of the enzyme, the amount of lysine assayed increases markedly (curve labelled 'boiled cells'). The difference in the quantities of lysine measured with the intact cells and with the disintegrated cells corresponds to the lysine liberated from the internal environment by disruption of the cell-wall, through which the enzyme cannot pass. Investigation of the amount of such lysine contained in the internal environment of the cells shows that its concentration depends upon that existing in the external environment, the ratio of the internal to external concentration varying from 2–3 for external concentrations of the order 0·02 M to approximately 20 for external concentrations of the order 0·001 M.

A Difference in Metabolic Requirements of Meningococcus and Gonococcus.

SummaryA rapid and heavy growth of meningococcus developed on an agar medium containing tryptic digest of casein, dextrose and buffer at pH 7.4-7.6. However, this medium failed to support growth of the gonococcus. Addition to the medium of a small amount of cystine, however, stimulated growth of this organism. The growth of gonococcus on this casein digest medium, with cystine, was as heavy as that of meningococcus. The growth of meningococcus did not seem to be improved by this addition of cystine to the medium.

Production of Small-Colony Variants of Staphylococcus aureus.

We have previously reported the production of small-colony variants, or G forms, after prolonged serial transfer of S. aureus in lithium chloride broth or plain broth media.1, 2 Swingle3 subsequently confirmed these results and also obtained these forms after extended growth in nutrient broth or casein-digest medium. Under these conditions the variants occur only rarely and in small numbers, seldom exceeding 1.0% of the colonies on a plate. Chinn4 in a study of the metabolic activity of the small colony variants of S. paradysenterio Sonne and S. aureus found them to be metabolically far less active than the normal forms, having a reduced generation-time, reduced fermentative powers, lessened ability to reduce methylene blue and a slower cataphoretic velocity. This lowered metabolic activity and the ability to revert to forms identical with the parent culture furnishes some evidence that small-colony variants may be forms whose metabolism has been greatly reduced by some unknown factor.Quastel and Woolridg...

The Relation of the Composition of the Culture Medium to the Formation of Endospores by Aerobic Bacilli

1. Spore formation in eight typical members of the genusBacillushas been studied.2. Three of these strains, including one species ofB. anthracis, have been found to be practically asporogenous under the experimental conditions. In general the following statements hold good for the sporogenous races studied.3. Spore formation is almost, or entirely, inhibited by cultivation on media rich in amino acids, such as tryptic digests of casein or meat. Similar inhibition results following cultivation on a medium containing reasonably high concentrations of a mixture of amino acids and asparagine.4. When such media are suitably diluted with standard inorganic salt solutions the percentage of spores formed is greatly increased, and frequently at least 99 per cent. of spores are formed if the dilution is sufficiently high.5. When simple nitrogenous compounds such as amino acids are added to a dilute casein digest medium in which sporulation is almost complete, a definite decrease in the percentage of spores present is observed. Asparagine, which is probably readily assimilated, apparently completely hinders spore formation in most cases. Other amino acids do not exert so pronounced an effect, and ammonium phosphate does not appreciably inhibit the formation of spores.6. The fact that the addition of glycine suppresses growth markedly when it is added to a dilute casein digest medium, but does not appreciably hinder sporulation, suggests that the formation of spores is not due to any toxic effect of added compounds, or compounds already present in the medium.7. Sporulation is almost complete in a “synthetic” medium in which low concentrations of ammonium phosphate and sucrose represent the sources of nitrogen and carbon, respectively. However, frequent transfers in such a medium may inhibit spore formation partially or entirely in certain instances. This effect probably depends upon the enhanced ability of the culture in question to utilise sucrose as a source of carbon when cultivated constantly in its presence.8. It is concluded, from the above data, that endospore formation in aerobic bacilli bears an inverse relationship to the amount of available nutrient material present in the culture medium.I am indebted to Prof. Sir F. G. Hopkins and Miss M. Stephenson for their constant encouragement during the progress of this work. My thanks are due to Mr Pirie of this Department who kindly furnished me with several of the amino acids employed, and to Dr Miles of the Department of Pathology for his kindness in supplying me with certain of the cultures.

Behaviour of Diphtheria Bacilli in Acidified Media, especially with reference to Virulence Tests

1. The growth of diphtheria bacilli in suger-free pure casein digest broth may be accelerated by the addition of dilute nitric, acetic and lactic acids, and delayed by certain monobasic fatty acids.2. The limiting acid zone for toxic and non-toxic varieties is unaffected by initial dilute acidifications by several acids, but incresaed by the addition of citric acid.3. The time periods and intensity of the reaction changes produced by diphtheria bacilli growing in a sugar-free casein digest medium are altered by the addition of certain acids. Toxic strains produce earlier and more progressive alkalinity when dilute acetic or nitric acid is included in the medium: and later and lessened alkalinity in the presence of butyric, formic, and propionic acids. Certain other typical acids do not exert any obvious action. Non-toxic strains are less affected by the selected acids, although butyric acid generally retards reaction changes.4. The time required for virulence testing is decreased, and the potency of the bacillus under examination is more evident when 1/200 η acetic or nitric acid is added to the culture medium. Acidified casein digest plus peptone broth yields practically the same results as acidified fermented veal peptone infusion. The acidification does not affect the metabolism of non-toxic strains.5. The disadvantages of unfermented meat infusions and the ascribed harmful effects of the fermentation products in fermented meat broths, may perhaps be avoided by the substitution of casein digest peptone medium. The advantages of the starting action of the glucose may be retained by adding acetic or nitric acid to the casein digest.6. Individual strains of diphtheria bacilli differ considerably in their capacity of altering the reaction of culture media, and in their toxic potency. It does not seem permissible to apply to newly isolated organisms all the published results obtained from cultures that have been repeatedly subcultured and established as toxin producers in nutrient bouillon. As a presumptive test in the virulence testing of bacilli just transferred from human tissues to artificial culture media the tendency of non-toxic strains to produce early formation of alkali in adequately buffered and adjusted unfermented veal broth may be thought worthy of extended trial.