Gari as culture medium for moulds

No significant differences (P<0.05 occurred in the frequency of isolation from soil of four common moulds – Rhizopus stolonifer, Monilia sitophila, Aspergillus niger and Penicillium sp., on malt extract agar (MEA) and gelled gari (carbohydrate gel). Growth of the moulds on slide microcultures of potato dextrose agar (PDA) and gari showed typical diagnostic features of the organisms, which were often clearer on gari. It is concluded that gari, which is much cheaper and far more readily available in Nigeria than MEA or PDA, is an effective alternative to the two standard mycological media.     

A new MSPQC for rapid growth and detection of Mycobacterium tuberculosis

Many of the deaths caused by tuberculosis (TB) in the world are due to wrong or late diagnosis. The World Health Organization (WHO) calls for better and cheaper TB tests method for this reason. In this paper, a new multi-channel series piezoelectric quartz crystal (MSPQC) sensor system was developed for rapid growth and detection of Mycobacterium tuberculosis. It is an automatic continuous monitoring system. The system was used to detect TB based on the volatile metabolic products NH(3) and CO(2) during the growth of M. tuberculosis. The metabolic products, diffusing from the medium into the KOH absorbing solution, resulted in the conductance change of the absorbing solution detected by the MSPQC sensitively. The frequency shift versus time response curves were recorded by self-developed software. Frequency detection time (FDT) corresponding to -100Hz in frequency shift value was used as a parameter to quantitatively determine M. tuberculosis H37Ra (an avirulent strain). H37Ra and 40 strains clinic positive samples were detected by the proposed system successfully. As for H37Ra, the FDT had a linear relationship with the logarithm of its initial concentration in the range of 10(2)-10(7) colony forming units (cfu)ml(-1) (R=-0.998) and the detection limit was low to 10cfuml(-1). 4% NaOH solution that can kill contaminating microorganisms and make M. tuberculosis alive was used as pretreatment reagent to provide selectivity to this method. Comparative tests were also carried out by using BACTECtrade mark MGITtrade mark 960 and conventional Lowenstein-Jensen (L-J) slants. The results showed that the proposed system was quicker than BACTECtrade mark MGITtrade mark 960 and it is also cheaper and will be widely used in TB tests in the world.     

A new growth medium for rapid selection and purification of Clostridium proteoclasticum transposon mutants

Clostridium proteoclasticum is commonly associated with the rumen microflora of pasture-fed cattle and sheep and has significant hemicellulose degradation abilities. Genes involved in plant fibre breakdown are commonly identified by producing site-specific mutations. However, the genetics of C. proteoclasticum and other closely-related Butyrivibrio/Pseudobutyrivibrio species is not well-established. Therefore random transposon mutants of C. proteoclasticum were generated by conjugation with Enterococcus faecalis containing Tn916. A new counter-selection agar medium was developed containing L-arabinose and D-raffinose as carbon sources, both of which are utilized by C. proteoclasticum only, and also ciprofloxacin, an antibiotic that suppresses the growth of E. faecalis. With this new medium the enhanced growth and more rapid separation of C. proteoclasticum transposon mutants from the background of E. faecalis cells was made, thereby facilitating the selection of transposon mutants and the identification of their Tn916 insertion sites.     

A selective medium for the rapid detection by an impedance technique of Pseudomonasspp. associated with poultry meat

A new medium for detecting and enumerating Pseudomonas spp. associated withpoultry meat spoilage by a rapid impedance technique was developed, after testing potentialgrowth promoters for eight Pseudomonas strains and inhibitors against eight competingstrains (Enterobacteriaceae) able to grow on the medium of Mead and Adams (1977). Four basalmedia (brain heart infusion, brucella broth, Shaedler broth and Whitley impedance broth (WIB))and a synthetic medium were evaluated. Whitley impedance broth was the best basal medium fordetecting variations in impedance in relation to Pseudomonas growth. The efficiency ofWIB was improved by adding compounds which enhanced the growth of Pseudomonas onthe synthetic medium. Among the incubation temperatures tested, 22°C proved to be the bestcompromise between growth of Pseudomonas associated with poultry meat spoilage andinhibition of competitors. Among the 15 inhibitory substances evaluated against Pseudomonas competitors, five were chosen for inclusion in the final medium : metronidazole,carbenicilline, cetrimide, cycloheximide and diamide (MCCCD medium). Preliminary resultsobtained from experiments with beef and pork meat showed that this medium could also be usedwithout diamide and at an incubation temperature of 25°C. The impedance technique usingMCCCD medium was then compared with an official method which uses the medium of Meadand Adams (1977) on 106 samples of poultry neck skin. The linear regression coefficient betweenthe two techniques was approximately r = 0·85. Impedance was able to detect 10³ Pseudomonas g⁻¹ within less than 19 h making it a promisingtechnique for predicting poultry meat spoilage.     

Development of a new culture medium for the rapid detection of Salmonella by indirect conductance measurements

The main difficulties in conductance medium development are to allow Salmonella to grow and produce a conductance signal while impeding growth of related species such as Escherichia coli and Citrobacter freundii . Various selective agents were screened for these capacities and a new medium was derived, named KIMAN (Whitley Impedance Broth basal medium supplemented with three selective components: novobiocin, malachite green and potassium iodide). This medium supported the growth of Salmonella serotypes and inhibited non-salmonella strains in pure cultures.     

A rapid coagglutination test for the detection and serogrouping of Legionellae

The applicability of coagglutination for the rapid detection and serogrouping of Legionellae has been investigated. The coagglutination reaction is carried out with the aid of self-made preparations of protein A containing staphylococci, sensitized with specific antibodies against the antigens of L. pneumophila (serogroups I to 6), L. bozemanii and L. micdadei. Preliminary heating of Legionella suspensions at 100% C for 15 min was used to prevent cross coagglutination reactions and ensure greater safety of laboratory personnel during the performance of the test. The results obtained demonstrate a high specificity of coagglutination. With the aid of the coagglutination reactions it has been shown that L. pneumophila strains isolated in Bulgaria belong to serogroup I. The coagglutination method is characterized by its rapidity, simplicity and feasibility. It is a useful and convenient means for the rapid detection and serogrouping of Legionellae.     

A rapid and sensitive method for the detection of histone peptides

Fluorescamine has been used to obtain a peptide map of a mixture of histones (H₃, H₂A, H₂B, and H₄) prepared from oocytes of Xenopus laevis. Fluorescamine was found to be more sensitive than o-phthalaldehyde or ninhydrin-Cd for the detection of peptide fragments obtained from tryptic digestion of oocyte histones of X. laevis and the peptic digestion of the β chain of insulin. Using the β chain of insulin for a comparison, the 8 major peptide fragments could be separated by electrophoresis within 30 min and were detectable at the picomols level. Some 70 peptide spots of X. laevis oocyte histones were resolved, thus permitting the analysis of this complex mixture of polypeptides without the need for prior separation.     

A selective-differential medium for detection of Streptococcus agalactiae

A practical culture medium which allows direct plating of milk samples for detection and differentiation of Streptococcus agalactiae within 48 hours is described. Most other micro-organisms likely to be present in these samples are inhibited. Although some strains of Staphylococcus species and ofStreptococcus faecalis are able to grow, they may be differentiated on the basis of reaction in the medium surrounding the colonies.     

A modified conductance medium for the detection of Salmonella spp

Selenite-cystine/trimethylamine oxide/dulcitol medium has been used in conjunction with conductance instruments to detect the presence of Salmonella spp. in foods and faeces. However, a small but significant number of salmonella strains were missed by this method. The majority of these strains were detected when dulcitol was substituted by mannitol and tested on two separate Malthus conductance instruments. Some strains of Citrobacter freundii and Escherichia coli continued to give false positive results. Attempts are made to explain why the substitution of mannitol for dulcitol gives an improved medium.     

A RAPD-PCR method for the rapid detection of Bacillus cereus

Distinction of Bacillus cereus from other closely related bacilli is challenging and new efficient methods are continually demanded. From our previous work on RAPD profiles of bacilli, we found a possibility that B. cereus strains could be distinguished from other bacilli. In this work, RAPD-PCR profiles of B. cereus strains were obtained using a 10-mer (S30) as a primer, and a B. cereus specific 0.91-kb band was produced from all tested strains. The RAPD-PCR procedure also successfully detected B. cereus from spiked cheonggukjang when B. cereus cells were present at more than 10(2)/g sample.     

A rapid chromatographic technique for the detection of dye-binding proteins

A rapid and inexpensive assay for dye-binding proteins has been developed. It depends on the separation of free and protein-bound sulfobromophthalein in 1-ml columns of Sephadex G-25 due to differential adsorption of the dye to the protein and to the Sephadex. With bovine serum albumin the calibration curve is linear between 0.03 and 3 mg of protein and is not affected by the presence of moderate concentrations of salt.     

Temperature and water activity minima for growth of spoilage moulds from meat

L owry , P.D. & G ill , C.O. 1984. Temperature and water activity minima for growth of spoilage moulds from meat. Journal of Applied Bacteriology 56 , 193–199.
Five species of fungi were isolated from mould spoilage on meat other than black spot. 'White spot' colonies yielded Chrysosporium pannorum or an Acremonium sp .; 'whiskers' colonies yielded Thamnidium elegans or Mucor racemosus , and blue-green colonies yielded Penicillium corylophilum. Chrysosporium pannorum was moderately xerotolerant with a minimum growth temperature of — 5C. The Acremonium sp. and P. corylophilum showed a similar level of xerotolerance but had a minimum growth temperature of — 2C. Mucor racemosus was no more xerotolerant than many spoilage bacteria and did not grow below - 1C, but grew rapidly at 3C and above. Thamnidium elegans grew at — 7C on supercooled medium and an intrinsic minimum growth temperature of — 10C was indicated. However, the low xerotolerance of this species precluded growth on frozen media below — 5C. It seems therefore that — 5C is the practical limiting temperature for mould growth on meat, and mould spoilage usually indicates that surfaces of freezer stored meats have approached and possibly exceeded 0C.     

Micromechanical oscillators as rapid biosensor for the detection of active growth of Escherichia coli

A rapid biosensor for the detection of bacterial growth was developed using micromechanical oscillators coated by common nutritive layers. The change in resonance frequency as a function of the increasing mass on a cantilever array forms the basis of the detection scheme. The sensor is able to detect active growth of Escherichia coli cells within 1 h which is significantly faster than any conventional plating method which requires at least 24 h. The growth of E. coli was confirmed by scanning electron microscopy. This new sensing method for the detection of active bacterial growth allows future applications in, e.g., rapid antibiotic susceptibility testing by adding antibiotics to the nutritive layer.     

Development of a defined medium supporting rapid growth for Deinococcus radiodurans and analysis of metabolic capacities

A morpholinepropanesulfonic acid (MOPS)-buffered rich defined medium (RDM) was optimized to support a reproducible 2.6-h doubling time at 35 °C for Deinococcus radiodurans R1 and used to gain insight into vitamin and carbon metabolism. D. radiodurans was shown to require biotin and niacin for growth in this medium. A glutamine–serine simple defined medium (SDM) was developed that supported a 4-h doubling time, and this medium was used to probe sulfur and methionine metabolism. Vitamin B₁₂ was shown to alleviate methionine auxotrophy, and under these conditions, sulfate was used as the sole sulfur source. Phenotypic characterization of a methionine synthase deletion mutant demonstrated that the B₁₂ alleviation of methionine auxotrophy was due to the necessity of the B₁₂-dependent methionine synthase in methionine biosynthesis. Growth on ammonium as the sole nitrogen source in the presence of vitamin B₁₂ was demonstrated, but it was not possible to achieve reproducibly good growth in the absence of at least one amino acid as a nitrogen source. Growth on sulfate, cysteine, and methionine as sulfur sources demonstrated the function of a complete sulfur recycling pathway in this strain. These studies have demonstrated that rapid growth of D. radiodurans R1 can be achieved in a MOPS-based medium solely containing a carbon source, salts, four vitamins, and two amino acids.     

A minimal medium for growth of Pasteurella multocida

Abstract We developed a minimal medium supporting the growth of both toxigenic and nontoxigenic strains of Pasteurella multocida to optical densities of > 0.5 (600 nm ). P. multocida P1059 (ATCC 15742), one of a number of strains which can cause fowl cholera, was used as the model strain in this study. The medium was composed of 17 ingredients including cysteine, glutamic acid, leucine, methionine, inorganic salts, nicotinamide, pantothenate, thiamine, and an energy source. Leucine was not required for growth but was stimulatory, and thiamine could be replaced by adenine. An additional 46 strains of P. multocida were tested, and 40 out of 46 (87%) strains grew as well as strain P1059 through a minimum of 10 serial transfers. P. multocida toxin (PMT) was produced when cells of a known toxigenic strain (P4261) were cultivated in the minimal medium. No growth of Pasteurella haemolytica or Pasteurella trehalosi strains was observed in this minimal medium.