Membranes with a pore size rating of 0.2μm are recommended for the sterilisation of liquids by filtration and are validated for this purpose by a retention test with Pseudomonas diminuta. Practices for retention testing were found to vary among the membrane manufacturers and only one type of commercial 0.2μm rated membrane was found to reliably retain P. diminuta. The retention for P. diminuta given by experimental grafted membranes was studied and was sometimes higher than that given by non-grafted membranes due to obstruction of the pores by graft material. The dimensions for individual cells of P. diminuta was studied by scanning electron microscopy and a rapid electronic method. Bacteria of larger dimensions than the pore size rating of experimental membranes were found in test permeates. It was shown that cells from an aerated P. diminuta culture were larger than cells from a similar but stationary culture. A retention test procedure for 0.2 μm rated membranes using cross-flow filtration was developed. The procedure simulated process conditions and enabled tubular ceramic monolithic membranes and flat-sheet membranes to be retention tested with P. diminuta. It is feasible that a standard retention test using cross-flow filtration can be developed. The time needed for results from current retention test procedures to become available is a consequence of using traditional cultural techniques for permeate analysis. Test procedure were developed using three popular methods for the rapid detection and enumeration of bacteria (ATP luminescence, impedance microbiology and the DEFT) for the detection and enumeration of P. diminuta in retention test permeates. The method using ATP luminescence was found to be the most applicable. The development of a bioluminescent strain of P. diminuta through genetic engineering will enable the rapid, sensitive and straightforward retention testing of 0.2 μm rated membranes. Retention tests using a bioluminescent strain of Escherichia coli containing the structural genes for bacterial luciferase indicated that the proposed test is feasible. Developments were made towards cloning the same genes into P. diminuta. The use of all bioluminescent micro-organisms for membrane retention testing is the subject of a patent application and a proposal for a three year SERC research grant.

Wood products all require treatment to mitigate phytosanitary risk prior to exportation. The most common phytosanitary treatment applied to Pinus radiata logs is Methyl Bromide (MeBr). The Environmental Protection Agency (EPA) in 2010 stated that MeBr must not be release into the atmosphere past 2020. This poses a problem for New Zealand log exports. Radiation has been identified as a possible alternative phytosanitary treatment for export wood products. This study aimed to quantify the effective dose of radiation necessary to sterilise two forest pest species; Arhopalus ferus and Hylurgus ligniperda. These species are representative of two different types of forestry pests; bark beetles (H. ligniperda) and wood borers (A. ferus). All applicable life stages for both species were tested. Arhopalus ferus adults were the most susceptible life stage identified with an LD99 of 30.2Gy ± 13.5 Gy (95% confidence interval). Arhopalus ferus eggs were less susceptible with a LD99 of 750Gy ± 776Gy observed; however there is low confidence in this result due to a methodological issue in one treatment replicate. Hylurgus ligniperda eggs were observed to be less susceptible than A. ferus eggs with a LD99 of 289Gy ± 92Gy. Results for the other life stages were inconclusive due to poor control survival, however the information gained was used to develop improved methods for further experimentation, which is on-going and showing positive results so far. The results of this experiment have indicated that radiation can be an effective method of sterilising forestry pests. To date radiation has not been used as phytosanitary risk mitigation for wood exports; however it is widely used for risk mitigation in agricultural products. Currently there remains a large amount of unknown information regarding, the effectiveness for irradiation of logs, the effective dose require for sterilisation of the most tolerant forestry pest and public acceptability of irradiation as a phytosanitary treatment. These knowledge gaps and an economic assessment must be completed before irradiation can be used as a phytosanitary risk mitigation technique for forestry products.

Univ., Diss.--München, 1994.

Zugl.: Hamburg, Univ., Diss., 2008

Současná pandemická situace ukázala obrovské dopady rozšíření infekce na společnost, i přesto stále ještě chybí jednotný přístup k problematice infekčního odpadu. Cílem této práce je představení metodiky nakládání s infekčním odpadem v rámci regionu v kontextu situace v Česku a Evropské Unii. V teoretické části práce je uveden legislativní rámec, charakter produkce, možnosti dekontaminace a podmínky spalování infekčního odpadu. Na základě těchto poznatků je navržena metodika pro nakládání s infekčním odpadem v rámci regionu. V praktické části práce je pak metodika aplikována na konkrétní region – Královéhradecký kraj. V rámci metodiky jsou navrženy možné scénáře nakládání s infekčním odpadem a vybrány nejvhodnější z nich z pohledu zdravotních rizik, rozpočtové zátěže a dopadu na životní prostředí. Na závěr je metodika kriticky zhodnocena a srovnána s metodikami jiných studií na podobné téma.

Doctor Legum - LLD / International standards recognise the basic right of all women and girls to make free choices about reproduction including the number if any, spacing and timing of their children without being subjected to discrimination, coercion, or violence. The enjoyment of this right by many women in the world has overtime been interfered with through forced sterilisation which has a salient history beginning with the eugenics movement in the 20th century indicating a disproportionate impact on the poor, ethnic minorities, women with disabilities, transgender group, as well as women living with HIV.

Konventionelle hitzesterilisierte, glukosehaltige Peritonealdialyselösungen (PDL) sind aufgrund ihres niedrigen pH-Wertes, ihrer hohen Glukosekonzentration und Osmolalität und ihres Gehaltes an Glukosedegradationsprodukten (GDP) bioinkompatibel. Alternativen zu glukosehaltigen PDL stellen aminosäuren- oder icodextrinhaltige PDL dar. Daneben enthalten auch neuere Glukose-PDL in Doppelkammersystemen aufgrund der Sterilisation von Glukose bei sehr niedrigem pH nur noch sehr geringe GDP-Konzentrationen. In dieser Arbeit wurden die akuten und chronischen Wirkungen verschiedener PDL auf humane peritoneale Mesothelzellen (HPMC) untersucht. Konfluente HPMC wurden mit den zu testenden PDL (glukosehaltige hitze- versus filtersterilisierte und konventionelle versus Doppelkammer-Glukose-PDL, 1% Aminosäuren-PDL und Icodextrin - alle bei neutralem pH-Wert) akut (1-4 Stunden Präinkubation) bzw. chronisch (bis zu 10 Tage) inkubiert. Die Zellvitalität (MTT-Assay) und IL-1beta-stimulierte Sekretion von IL-6 (Zellfunktion) wurden untersucht. Die akute und chronische Exposition von HPMC gegenüber hitzesteriliserten Peritonealdialyselösungen führte zu einer signifkanten Reduktion von Vitalität und Funktion der Zellen. Demgegenüber führte die Inkubation mit filtersteriliserten PDL und GDP-armen PDL zu einer weniger starken Beeinflussung von Vitalität und Funktion. Die aminosäurenhaltige PDL beeinflusste weder akut noch chronisch die Vitalität bzw. Funktion der Zellen negativ. Hingegen unterschied sich die icodextrinhaltige Lösung nicht wesentlich von der hitzesteriliserten PDL mit hohem Glukoseanteil. Die Verringerung des Gehaltes an GDP in PDL mittels Filtersterilisation bzw. alternativer Sterilisation in Zweikammerbeuteln hat einen positiven Einfluss auf Vitalität und Funktion von HPMC in vitro. Der Ersatz des osmotischen Agenz hingegen bedeutet nicht zwansgläufig eine bessere Biokompatibilität. / Conventional heat-sterilized glucose-containing peritoneal dialysis fluids (PDF) are bioincompatible due to their acidic pH, high glucose concentration and resulting hyperosmolality, and the presence of glucose degradation products (GDP). Alternatives to these solutions are PDL containing amino acids or icodextrin as the osmotic agent. Furthermore, novel glucose-based PDF contain only trace amounts of GDPs due to the sterilisation of glucose at very low pH in a dual-chamber container system. The present study examines the acute and chronic effects of different PDL on human peritoneal mesothelial cells (HPMC). Confluent HPMC were exposed to the different test PDF (glucose containing heat- versus filter-sterilised PDF and conventional versus dual-chambered glucose PDF, 1% amino-acid PDF and icodextrin – all at neutral pH) in an acute (1-4 hours preincubation) and chronic (up to 10 days) cell culture model. Cell viability (MTT assay) and IL-1beta stimulated IL-6 (cell function) release were assessed. Acute and chronic exposure of HPMC to heat-sterilised PDF resulted in a significant reduction of viability and cell function. In contrast, the incubation of filter-sterilised PDF and dual chambered low GDP solution had only minor effects on cell viability and function. Neither viability nor cell function were negatively affected by the amino-acid PDF following acute and chronic exposure. However, incubation with icodextrin resulted in a similar degree of inhibition as compared to incubation with conventional heat-sterilised glucose PDF. In conclusion, removal of GDP from PDF either via filter-sterilisation or manufacture in dual chambered containers helps to conserve viability and function of HPMC in vitro. However, the replacement of the osmotic agent per se does not necessarily result in improved biocompatibility.

Ziel dieser Arbeit war es, die Morphologie und mechanischen Eigenschaften von einzelnen nativen Typ-I-Kollagenfibrillen bei verschiedenen Umgebungsbedingungen zu untersuchen. Um auf der Nanometerskala die mechanischen Eigenschaften der Fibrillenoberfläche und des Fibrillenvolumens bestimmen zu können, wurde die dynamische (MUSIC-Mode) und statische (FD) Rasterkraftmikroskopie (engl.: Atomic-Force-Microscopy – AFM) genutzt. Die dynamische Spektroskopie erlaubte Rückschlüsse auf die Änderung der mechanischen Eigenschaften von Oberflächen (unter Einbeziehung der ersten 10 nm) bei Variation der Umgebungsbedingungen, wohingegen die statische Kraftspektroskopie Einblicke in das mechanische Verhalten der Volumenphase des Materials (Bulk) gestattete. Es wurden einzelne ovine (vom Schaf stammende) Kollagenfibrillen hinsichtlich ihrer mechanischen und morphologischen Veränderung als Folge von variierten Umwelteinflüssen (Salzkonzentrations-änderung des umgebenden Mediums sowie beim Durchlaufen des Peroxo-essigsäure/Ethanol-Sterilisationsprozesses) untersucht. Im Einzelnen konnte mit dem MUSIC-Mode-AFM und der FD-Spektroskopie die Denaturierung einer Kollagenfibrille in Abhängigkeit der Salzkonzentration beobachtet werden. Ebenfalls konnte mittels der MUSIC-Mode und der FD-Spektroskopie für mehrere Fibrillen unterschiedlichen Durchmessers nachgewiesen werden, dass während des Entfettungsschrittes des Peroxoessigsäure/Ethanol-Sterilisationsprozesses irreversible Veränderungen ihres Quellverhaltens sowie ihrer mechanischen Eigenschaften auftreten.:1 Einleitung ...................................................................................... 8 2 Zielstellungen dieser Arbeit .......................................................... 9 3 Kollagen ...................................................................................... 10 3.1 Aufbau und Struktur .......................................................................... 10 3.2 Die Rolle des Wassers und der Einfluss des Salzes auf die mechanischen Eigenschaften von Kollagen .......................................... 14 3.3 Einfluss der Sterilisation auf Kollagen ................................................ 20 3.4 Bisherige Untersuchungen von Kollagen mit dem AFM – Stand der Technik .................................................................. 24 4 Methoden .................................................................................... 29 4.1 Rasterkraftmikroskop (Atomic Force Microscope–AFM) .................... 29 4.1.1 Statische Kraftspektroskopie ........................................................... 33 4.1.2 Dynamische Kraftspektroskopie ...................................................... 35 4.1.3 Kontaktmodelle ............................................................................... 39 4.1.4 Spitze-Probe Wechselwirkungen ...................................................... 41 4.2 Infrarotspektroskopie [152] .................................................................. 48 4.2.1 Funktionsweise des ATR-FTIR ....................................................... 50 4.2.2 IR-Spektroskopie an Proteinen ........................................................ 51 5 Experimenteller Teil ................................................................... 53 5.1 Herstellung der Kollagenproben .......................................................... 53 5.2 Durchführung der AFM-Messungen .................................................... 53 5.2.1 Durchführung der AFM-Messung zum Kapitel „Einfluss der NaCl-Konzentration auf Typ-I-Kollagen“ ................................................. 54 5.2.2 Durchführung der Untersuchungen zum Sterilisationsprozess ......... 55

Xenos Ltd. is a technology driven food company, that specializes in aseptic processing and packaging beverage products in bottles. Their aseptic filling technology is based on packaging sterilization with combined treatments of oxidizing agents and Ultraviolet radiation. Recent research studies have suggested that there is a synergistic effect of hydrogen peroxide (0.5 – 1 %) plus UV on inactivation of microorganisms including spores. Advantages of the combined treatment include rapid inactivation, minimum hydrogen peroxide residue in products, with the method being applicable to a wide range of packaging types. Based on this principle, a unique aseptic packaging technique has been developed by Xenos Ltd., which utilizes the combination of vaporized Perform (a commercial sterilizing agent manufactured by Orica Chemnet containing 25% hydrogen peroxide and 5% peracetic acid) and UV radiation at 7.5 – 12.5 W/m2. The aim of the project was to improve and validate the effectiveness of the packaging sterilization process through challenge tests. Challenge tests were conducted using Bacillus subtilis spores as the test microorganism to determine the log reductions delivered by the packaging sterilization system. The tests were firstly carried out on a pilot plant scale aseptic filling machine, in order to test the sterility of the small scale system, and investigate processing parameters (operational conditions) which could affect and improve sterility. The established operational conditions for achieving target sterility were used for designing and modifying an upgrade aseptic packaging system. Finally validation of the upgrade packaging sterilization system was conducted through challenge tests to prove sterility. It is highly recommended that in order to ensure sterility, the packaging sterilization system with vaporized Perform plus UV treatment must meet the requirements listed below during the sterilization process:  Hydrogen peroxide concentration of Perform condensate on bottles (after steaming) is best within 0.5 – 1 %;  Perform loading level should be minimum 300 mg/bottle after vaporized Perform treatment;  UV treatment time applied is greater than 2 seconds during UV treatment;  At least 20 seconds of penetration time (time between Perform treatment and UV treatment) should be allowed. The upgrade sterilization system used by Xenos Ltd. has been improved to meet the above operational conditions. With spore loading level of 106 per bottle and 105 per cap, the system is able to deliver at least a 6 log reduction of B. subtilis spores on PET or glass bottles and a 5 log reduction on bottle caps. Moruzzi et al. (2000) stated that at least a 4 log reduction is commercially required for an aseptic packaging process. Therefore, the system’s sterility would meet the commercial acceptable sterility.

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