Use of a blocking primer allows selective amplification of bacterial DNA from microalgae cultures

The production of algae is a crucial component of many aquaculture systems and the role of bacteria in this process is an important although complex one. We report the development of a new blocking primer that allowed PCR amplification of bacterial DNA in the presence of algal chloroplast DNA.     

Development of the optimal inoculation conditions for microcarrier cultures

The environmental conditions under which anchorage-dependent mammalian cells are grown are not necessarily those under which a culture should be initiated. Cell attachment is a physical process, and those factors which affect forces involved in cell attachment differ from the biological factors which affect cell growth. We have conducted an extensive experimental study to define clearly the optimal environmental conditions for MRC-5 cell attachment onto microcarriers. These inoculation conditions are particularly important when the serial propagation of mammalian cells on microcarriers is considered as in a human vaccine production process. The conditions which were investigated are: initial serum content (% v/v), initial pH, inoculation level (cells/bead), agitation rate (rpm), and the concentration of microcarriers (g/L). The initial distribution of attached cells was found to have a significant affect on the overall efficiency of anchorage-dependent cell cultures, and was used to evaluate attachment efficiency. Based on the experimental results, we propose an optimized protocol for the inoculation of microcarrier cultures.     

Effect of low inoculation density in the scale-up of insect cell cultures

The scale-up of insect cell cultures and the production of baculovirus with these cultures is dependent on the inoculation density applied. The effect of applying a low inoculation on the specific growth rate and on the duration of the lag phase was tested. Three different cell lines, HzAm1, Ha2302, and Sf21 were tested in a total of five cell line/medium combinations. Growth in suspension culture was examined, and data obtained were fitted with the Gompertz equation. A significant decline in specific growth rate with decreasing inoculation density was observed in all cell line/medium combinations, except for HzAm1. No critical inoculation density, below which no growth would occur, was found. In suspension culture in shake flasks, an inoculation density of 5 x 10(4) cells/mL is achievable, without severely influencing the overall growth rate. A lower inoculation density in suspension culture results in less steps in the scale-up process and might be a tool in bypassing the viral passage effect.     

Use of hen lysozyme for protection against bacterial contamination ofin vitro embryo cultures

Summary Curative treatments with antibiotics and hen egg white lysozyme (HEWL) were used to salvage embryo cultures contaminated withBaccillus subtilis. The use of HEWL gave good control ofBaccillus subtilis, but no control ofErwinia. HEWL was better than antibiotics, being much less phytotoxic. The antibiotics piperacillin, ampicillin and imipenem were also found to be ineffective againstErwinia. HEWL, at a final concentration of 1 mg per mL, was used as a preventive and curative agent for routine use in embryo culture ofTriticum aestivum and other Triticeae, as it cured from 30% to 50% of bacterial contamination problems over a one year period. Standardin vitro culture precautions remained essential, as certain bacteria were not controlled by HEWL.     

Comparison of the effect of bacterial inoculation in musculocutaneous and fasciocutaneous flaps

The skin fascial flap is now recognized as a reliable flap for use in reconstructive surgery. The fasciocutaneous flap has been advocated for coverage of chronic infected wounds after debridement as an alternative to the musculocutaneous flap. Previous experimental and clinical studies have demonstrated the superior resistance of the musculocutaneous flap as compared to the random-pattern flap to bacterial inoculation. A canine model is presented for comparison of the effect of bacterial inoculation in fasciocutaneous and musculocutaneous flaps of similar dimensions. The area of skin necrosis secondary to bacterial inoculation was similar in these two flap types despite greater blood flow and skin oxygen in the fasciocutaneous flap. In a study of closed wound spaces formed by the deep surface of these two flap types, a greater degree of inhibition and elimination of bacterial growth and more collagen deposition are observed in the musculocutaneous wound space than in the fasciocutaneous flap.     

Equipment for the automatic aseptic inoculation and sampling of microbial cultures

Equipment has been designed and constructed for the automatic aseptic inoculation and sampling of microbial cultures. It enables fermentations to be started and samples to be taken round-the-clock, according to a predetermined program. The equipment is suitable for microbial processes on both a laboratory and a factory scale.     

The proportion of mutants in bacterial cultures

The proportion of mutants in a growing culture of organisms will depend upon (a) the rate at which the wild cells produce them (with or without growth), (b) the back mutation rate, and (c) the growth rates of the wild and mutant cells. If the mutation rate without growth and the back mutation rate are neglected, the growth of a mutant is expressed by See PDF for Equation and the ratio of the mutant to wild by See PDF for Equation in which λ = mutation frequency rate constant, "mutation rate," A = growth rate constant of wild cells W, B = growth rate constant of mutant cells M. If the term [B – (1 – 2λ)A] is positive, the proportion of mutants increases continuously. If it is negative, the proportion of mutants reaches a constant value See PDF for Equation If mutation is assumed to occur without growth at the rate C, then the corresponding equations are (11), (12), and (14). See PDF for Equation If (B + C – A) is negative and t = ∞, See PDF for Equation If C << A, See PDF for Equation     

Decomposition of the herbicide bromoxynil in soil and in bacterial cultures

It was found in field, and laboratory experiments that of 50 ppm of the herbicide bromoxynil (3,5-dibromo-4-hydroxybenzonitrile added to grey forest soil 20-80% were still detected after three months). Bromoxynil did not influence (except for a short-termed stimulation of the number of bacteria) the amount and composition of the basic groups of soil microorganisms. In enrichment cultures of soil microorganisms metabolie products of bromoxynil decomposition (3,5-dibromo-4-hydroxybenzamide and 3,5-dibromo-4-hydroxybenzoic acid) were detected and a stimulating effect of cosubstratos on its decomposition was demonstrated. Bromoxynil concentration, aeration conditions and the presence of cosubstrates (ribose in particular) influenced the rate and degree of the decomposition process inPsevdomonas putida. In addition to the degradation products mentioned above, production of methoxylated and partially dehalogenated aromatic compounds was detected.