Development of a new method for the characterisation of bioreactors concerning heat and mass transfer

Since the specific heat transfer coefficient (UA) and the volumetric mass transfer coefficient (kLa) play an important role for the design of biotechnological processes, different techniques were developed in the past for the determination of these parameters. However, these approaches often use imprecise dynamic methods for the description of stationary processes and are limited towards scale and geometry of the bioreactor. Therefore, the aim of this thesis was to develop a new method, which overcomes these restrictions. This new approach is based on a permanent production of heat and oxygen by the constant decomposition of hydrogen peroxide in continuous mode. Since the degradation of H2O2 at standard conditions only takes place by the support of a catalyst, different candidates were investigated for their potential (regarding safety issues and reaction kinetic). Manganese-(IV)-oxide was found to be suitable. To compensate the inactivation of MnO2, a continuous process with repeated feeds of fresh MnO2 was established. Subsequently, a scale-up was successfully carried out from 100 mL to a 5 litre glass bioreactor (UniVessel®)To show the applicability of this new method for the characterisation of bioreactors, it was compared with common approaches. With the newly established technique as well as with a conventional procedure, which is based on an electrical heat source, specific heat transfer coefficients were measured in the range of 17.1 – 24.8 W/K for power inputs of about 50 – 70 W/L. However, a first proof of concept regarding the mass transfer showed no constant kLa for different dilution rates up to 0.04 h-1.Based on this, consecutive studies concerning the mass transfer should be made with higher volume flows, due to more even inflow rates. In addition, further experiments are advisable, to analyse the heat transfer in single-use bioreactors and in larger common systems.

Comparasion of polyacrylamide gel electrophoresis (PAGE), immuno-electron microscopy (IEM) and enzyme immunoassay (EIA) for the rapid diagnosis of rotavirus infection in children

Detection of rotavirus RNA by polyacrylamide gel electrophoresis (PAGE) proved to be a highly sensitive and rapid diagnostic test. A comparison of this assay with immuno-electron microscopy (IEM) and enzyme immunoassay (EIA) in 245 faeces from children with gastroenteritis revealed complete agreement between the three assays in 238 (97.14%) samples. Among 75 samples positive in at least one of the three assays, negative results were observed in 5 (6.48%) by PAGE, in 6 (6.76%) by EIA and in none by IEM. Silver staining greatly increased the sensitivity of the PAGE assay. We conclude that although IEM remains the most sensitive and rapid rotavirus diagnostic assay, the PAGE technique has many advantages in its favour, including the non-requirement of expensive equipment, the use of only chemically defined reagents and the capacity to distinguish virus subgroup and variants and to detect non-crossreactive rotaviruses which are missed in serological assays.

Numerical simulation of combined heat transfer phenomena (conduction, convection, and radiation). Application to he optimisation of thermal systems

Thermal systems interchanging heat and mass by conduction, convection, radiation (solar and thermal ) occur in many engineering applications like energy storage by solar collectors, window glazing in buildings, refrigeration of plastic moulds, air handling units etc. Often these thermal systems are composed of various elements for example a building with wall, windows, rooms, etc. It would be of particular interest to have a modular thermal system which is formed by connecting different modules for the elements, flexibility to use and change models for individual elements, add or remove elements without changing the entire code. A numerical approach to handle the heat transfer and fluid flow in such systems helps in saving the full scale experiment time, cost and also aids optimisation of parameters of the system. In subsequent sections are presented a short summary of the work done until now on the orientation of the thesis in the field of numerical methods for heat transfer and fluid flow applications, the work in process and the future work.

Second contribution to the knowledge of the larvae of the Raviniini (Diptera, Sarcophagidae) based on observations using scanning electron microscope

First and second instar larvae of some Sarcophagidae (Diptera) of the tribe Raviniini are described on observations with a scanning electron microscope.

Third contribution to the knowledge of the Raviniini (Diptera, Sarcophagidae), based on observations of the larvae, using scanning electron microscope

L1, L2 and L3 of Oxysarcodexia paulistanensis (Mattos), L3 of O. confusa Lopes, L2 of Ravinia belforti (Prado & Fonseca) and L2 of Oxyvinia excisa (Lopes) were described and figured using scanning electron microscope.

Distal dendrites of frog motor neurons: a computer-aided electron microscopic study of cobalt-filled cells.

With the aid of the cobalt labelling technique, frog spinal cord motor neuron dendrites of the subpial dendritic plexus have been identified in serial electron micrographs. Computer reconstructions of various lengths (2.5-9.8 micron) of dendritic segments showed the contours of these dendrites to be highly irregular, and to present many thorn-like projections 0.4-1.8 micron long. Number, size and distribution of synaptic contacts were also determined. Almost half of the synapses occurred at the origins of the thorns and these synapses had the largest contact areas. Only 8 out of 54 synapses analysed were found on thorns and these were the smallest. For the total length of reconstructed dendrites there was, on average, one synapse per 1.2 micron, while 4.4% of the total dendritic surface was covered with synaptic contacts. The functional significance of these distal dendrites and their capacity to influence the soma membrane potential is discussed.

Role of a mouse monoclonal IgE antibody in passive transfer of immunity to Schistosoma japonicum infection

We have been able to produce a mouse monoclonal IgE antibody specific to an adult worm antigen extracted from Schistosoma japonicum (Sj). The antibody was able to elicit passive cutaneous anaphylaxis in the rat skin against Sj with the highest titer of 1:256,000 but did not cross-react with S. mansoni antigen. The antibody recognized a 97-kDa molecule expressed on the surface of mechanically transformed schistosoma of S. japonicum. Passive transfer of the antibody into mice in the early stage of challenge infection resulted in a partial but significant reduction of recovery of adult worms. Induction of eosinophilia by an oral administration of embryonated eggs of Toxocara canis prior to challenge infection enhanced the reduction.

Rapid detection by transmission electron microscopy of mycoplasma contamination in sera and cell cultures

Transmission electon microscopy has been employed for the rapid detection of mycoplasma in sera and cell cultures. High speed centrifugation of sera or low speed centrifugation of cell debris, followed by negative staining of the resuspended pellet, detected mycoplasma contamination more frequently than a culture method followed by direct fluorescence (DAPI), which was used as a control procedure. The appearance of the mycoplasma cell border and content gives some information about particle viability.

Food availability affects the maternal transfer of androgens and antibodies into eggs of a colonial seabird.

Mothers can improve the quality of their offspring by increasing the level of certain components in their eggs. To examine whether or not mothers increase deposition of such components in eggs as a function of food availability, we food-supplemented black-legged kittiwake females (Rissa tridactyla) before and during egg laying and compared deposition of androgens and antibodies into eggs of first and experimentally induced replacement clutches. Food-supplemented females transferred lower amounts of androgens and antibodies into eggs of induced replacement clutches than did non-food-supplemented mothers, whereas first clutches presented no differences between treatments. Our results suggest that when females are in lower condition, they transfer more androgens and antibodies into eggs to facilitate chick development despite potential long-term costs for juveniles. Females in prime condition may avoid these potential long-term costs because they can provide their chicks with more and higher quality resources.

Scanning electron microscopy of the egg and the first instar larva of Dermatobia hominis (Diptera, cuterebridae)

The egg and the first instar larva of Dermatobia hominis were described based on observation with a scanning electron microscope.

Morphological evidence by scanning electron microscopy of excretion of metacyclic forms of Trypanosoma cruzi in vector's urine

Comparision by scanning electron microscopy (SEM) of Trypanosoma cruzi flagellates attached to the cuticle of the rectal gland of infected Dipetalogaster maxima nymphs, showed marked differences before amd after feeding. Before feeding numerous metacyclic trypomastigotes were observed among the abundant epimastigotes that formed the carpet of flagellates. On the other hand, in insects that were allowed to urinate for 24 hours after a meal, the metacyclics were scarce,indicating that they had been detached by the urine flow. An asymetric type of cell division, probably originating both an epi-and a trypomastigote, was occasionally observed. The occurrence of swellings at different levels of the flagella of epimastigotes suggests that secondary sites of attachment may be common.