Simple technologies are required for reducing the post harvest losses of horticultural produce. Edible films are being studied extensively for application on fresh and cut fruits and vegetables. Tomato, being a very nutritious and important food and a highly perishable climacteric fruit, this study was planned to investigate the application of chitosan films. Chitosan is a biodegradable waste product from sea food and is safe for consumption. With a view to broaden its antimicrobial activity it was combined with lysozyme, a lytic enzyme. Since the edible films are sensitive to changes in temperature and humidity, they were studied under ambient and optimal conditions of storage. / This study showed that 1% chitosan was more suitable for tomatoes for storage at both conditions of ambient and low temperature. Respiration study showed that 1% chitosan treatments resulted in more favorable levels of CO 2 production and internal O2. This was reflected in the quality of tomatoes held under these conditions. Two per cent chitosan films were unsuitable due to their high CO2 production and low internal O 2 levels. Spoilage was more apparent in this treatment. Lysozyme addition did not show any additional benefit. / The research in this study has demonstrated that the selection of edible films for horticultural produce needs to be integrated with the requirement of storage conditions of the produce.

Initial experiments were conducted to determine the microbial development in blocks of ground beef. In the top layers of the meat, Pseudomonas spp. predominated. Attempts were made, as well, to measure the microenvironmental changes occurring in ground beef using pH, oxygen, and redox potential micro-electrodes. / Some of the extrinsic (relative humidity, O(,2)), as well as intrinsic factors (pH, redox potential, ions, nutrients) affecting microbial development in aerobically stored meats, were examined. The decrease in glucose levels observed during the aerobic storage of longissimus dorsi (l. dorsi) muscle at 4(DEGREES)C, was accompanied by an increase in the activity of glucose dehydrogenase, and by the appearance of gluconate and 2-ketogluconate on the meat surface. / The attachment of various meat spoilage organisms to l. dorsi muscle was studied. Generally, the pseudomonads attached in higher numbers than the other bacteria, but possessed lower attachment strengths. Investigations on the attachment of two competing organisms to l. dorsi muscle demonstrated that limited competition occurs between the organisms. / A model of the microbial ecology of aerobically stored fresh beef was developed, based on existing evidence. / Various chemicals were added to minced or whole meat to evaluate their effectiveness as food preservatives. As a single compound, potassium sorbate appeared to have the greatest potential as a meat preservative. Meats > into solutions of 5.0% potassium sorbate for 1 min had their shelf-life substantially increased as compared with control samples > into water.

The technical feasibility of ohmic heating was evaluated for viscous liquids in static ohmic heating cells in three stages. First, electrical conductivities and time/temperature profiles were measured and compared for selected hydrocolloids (carrageenan, gelatin, pectin, starch and xanthan) in water at various concentrations (1--6%). Of the thickening agents examined, carrageenan gave the highest value for electrical conductivity and the shortest time to raise the temperature from 20 to 100°C. It was followed by xanthan. Pectin and gelatin samples were found to exhibit lower, but similar electrical conductivities and heating profiles. Starch samples had the lowest electrical conductivity and the temperature of starch solutions never exceeded 62°C within the specified time limit of 10000 s. / In the second stage, the effect of salt and acid concentration was evaluated on electrical conductivities and time/temperature profiles of the above selected hydrocolloids in water solutions at a concentration necessary to achieve a similar apparent viscosity of 0.2 Pa.s at 300 s-1 and 20°C (carrageenan 1.7%, pectin 2.5%, starch 4.3% and xanthan 2%). / In the third stage, measurements of electrical conductivities, time/temperature and heating rate profiles were performed applying six voltage gradients (5.26, 7.14, 13.16, 17.86, 21.05 and 25 V/cm), with two electrode cross-sectional surface areas (19.84 and 37.95 cm2) and the electrodes separated at three specific distances (10.05, 14.33 and 20.01 cm) at 150 V. Applied voltage gradients had a major effect on temperature and heating rate profiles but no effect on electrical conductivities. / Rheological properties of carrageenan, pectin, starch and xanthan solutions were investigated at various temperatures (20--80°C) and concentrations (carrageenan 1.5, 1.7 and 1.9%; pectin 2.3, 2.5 and 2.7%; starch 3.8, 4 and 4.2%; xanthan 1.6, 1.8 and 2%) in the presence of 1% salt. Both temperature and concentration influenced the rheological properties of solutions. / Finally, electrical conductivity measurements and rheological properties of starch at 4% and 1% salt were used as input to evaluate a theoretical model for the electrical, thermal and flow behaviour in a continuous ohmic heating unit. (Abstract shortened by UMI.)

The effects of irradiation dose (0, 0.5 and 1.0 kGy), various gas atmospheres and storage temperature (5, 15 and 25$ sp circ$C) on the physical, chemical, microbiological, and organoleptic changes in fresh pork were studied using factorial design experiments. The effects on toxin production by Clostridium botulinum were also investigated using challenge studies. Shelf life could be extended to 21 d when product was packaged in 0% O$ sb2$, irradiated at 1.0 kGy and stored at 5$ sp circ$C compared to 4 d for control samples. While the presence of O$ sb2$ in the package headspace enhanced the antimicrobial effects of low-dose irradiation, it adversely affected the organoleptic qualities of pork. Botulinum toxin was detected after only 2 d in all inoculated treatments stored at 25$ sp circ$C. At 15$ sp circ$C, toxin was produced faster when pork was initially packaged with O$ sb2$ or low levels of CO$ sb2$ (15-30%) as compared to 100% N$ sb2$. Higher levels of CO$ sb2$ (45-75%) delayed toxin production. In most treatments, spoilage preceded toxigenesis. Models were developed relating the above factors to the time until toxin production and to the probability of toxigenesis. Temperature, initial O$ sb2$ and irradiation were all significant factors.

High pressure (HP) processing, a novel technology, has excellent potential for non-thermal preservation of apple juice, the largest consumed fruit juice in North America. The objective of this research was to evaluate the application of HP treatment for inactivation of enzymes and destruction of microorganisms in apple juice. HP inactivation kinetics of selected enzymes (amylase, pectin methyl esterase and polyphenol oxidase) and microorganisms [Leuconostoc mesenteroides, Pichia membranaefaciens and Zygosaccharomyces badii, Escherichia coli (29055) and Escherichia coli (O157:H7)] in apple juice were evaluated under various test conditions (100--400 MPa, 0--60 min and 6--40°C) using a central composite design of experiments. The enzymes selected were of importance in apple juice preparation and/or storage stability of the processed juice. Microorganisms included those that are responsible for spoilage and/or public health concern as well as those that are indicative of unsanitary handling conditions. / Enzyme inactivation and microbial destruction due to pressure followed a dual-effect model consisting of a pressure pulse effect (PE) and a subsequent semi-logarithmic (first order) inactivation during the pressure hold-time. In general, results showed that inactivation of enzymes and destruction of microorganisms was more prominent at higher-pressure levels, higher temperature and longer treatment times, and at lower pH levels of juice. Pressure pulse effect was dependent on pressure level, with higher PE achieved at higher pressures. During the pressure-hold, as expected, the associated decimal reduction times (D values) decreased with an increase in pressure. Pressure dependency of D values was well described by the conventional death time model. The pressure resistance of enzymes and microorganisms varied, but complete inactivation of enzymes and destruction of microorganisms was possible with the combination of lower pH, higher pressure and higher temperatures. / Commercial PME from a citrus source was more pressure sensitive than PME from microbial source. Spoilage bacteria (L. mesenteroides) were more pressure resistant than the yeasts. E. coli enumerated on an enrichment media (supporting both injured and healthy cells) showed larger survivors and a greater resistance than on a more selective media. An increasing number of cells got injured than killed with the application of pressure treatment until they were all finally injured or killed. High-pressure treatment (pulse at 400 MPa or by holding about 10 min at 350 MPa and 30°C) resulted in complete destruction of the pathogenic microorganism E. coli (O157:H7) ensuring the public health safety of the process.

Listeria monocytogenes is a pathogenic, psychrotrophic microorganism that is ubiquitous in nature. L. monocytogenes has been isolated from numerous meat products, both fresh and processed, the incidence of contamination varying greatly. The ability of Listeria to grow in meats depends on temperature, pH, water activity (a$ sb{ rm w}$), nutrients, species and numbers of competing microorganisms, gaseous conditions, and levels of additional barriers. Therefore, methods to control the growth of L.monocytogenes are of great importance to food processors since this organism can grow under a wide range of environmental and storage conditions. Two methods of control, in conjunction with temperature, were studied in this project: (i) modified atmosphere packaging (MAP) and (ii) chitosan, to determine the optimum levels of these "hurdles" needed to effectively control the outgrowth of L.monocytogenes in both model broth and agar systems and in fresh pork loin. On the basis of these preliminary studies, a combination of chitosan as a dipping solution and modified atmosphere packaging were investigated to control the growth of L.monocytogenes in fresh pork loin. Pork loin samples were dipped in a 0.2% chitosan solution for 60 seconds and packaged under various atmospheres in Cryovac bags and stored at 5, 10 and 15$ sp circ$C up to 28 days. Samples were monitored for physical, chemical and microbiological changes throughout the storage period. Optimum control over the growth of L. monocytogenes was achieved using a combination of 100%N$ sb2$ + an Ageless FX oxygen absorbent and dipping in a 0.2% chitosan solution. Based on these studies, a combination of 0.2% chitosan and MAP could be used to extend the shelf life of pork without adversely affecting color, odor and exudate loss while inhibiting the growth of the pathogenic microorganism, L.monocytogenes. (Abstract shortened by UMI.)

In this research, initial studies were done to determine the effect of various methods of presentation involving chemical preservatives, water activity ($ rm a sb{w}$), and modified atmosphere packaging (MAP) on mold growth in an agar model system. Results showed that preservatives could completely inhibit mold growth for 2-40d depending on concentration and pH used. Gas packaging (60% or 80% CO$ sb2$), oxygen absorbents, alone or in combination with potassium sorbate, could also inhibit mold growth for $>$40d at ambient storage temperature using a Response Surface Methodology (RSM) approach. / The effects of various methods of applying potassium sorbate into pizza crusts via direct incorporation into the batter, surface spraying, and impregnation of packaging material with potassium sorbate to control mold spoilage of pizza crusts were also investigated. Results showed that the antimicrobial effect of potassium sorbate was negligible when the packaging material was impregnated with the inhibitor but more pronounced when it was incorporated directly into the dough or sprayed onto the product's surface. The inhibitory effect of potassium sorbate increased as both the pH and the inoculum level decreased. / Shelf life studies using low concentrations of potassium sorbate (1000 and 2000 p.p.m.) and MAP, alone and in combination with each other, showed that potassium sorbate, gas packaging or oxygen absorbents (Ageless FX) could extend the shelf life of pizza crusts and decrease the growth rate of molds, bacteria and yeast. Furthermore, when pizza crusts were packaged in 60% CO$ sb2$ or with an oxygen absorbent, in combination with potassium sorbate (1000-2000 p.p.m.), a shelf life of 42d was possible without compromising the sensory shelf life of the product. (Abstract shortened by UMI.)

<p><font face="CMR12"> <p align="left">This thesis deals with analysis and implementation of a positivity preserving numerical method for a vaccination model for the transmission dynamics of two HIVsubtypesnin a given community. The continuous model is analyzed for stability and equilibria. The qualitative information thus obtained is used while designing numerical method(s). Three numerical methods, namely, Implicit Finite Difference Method (IFDM), Non-standard Finite Difference Method (NSFDM) and the Runge-Kutta method of order four (RK4), are designed and implemented. Extensive numerical simulation are carried out to justify theoretical outcomes.</p> </font></p>

Three questions are considered in the context of the possible effects of seedassociated mycoflora, typified by Fusarium moniliforme, during hydrated storage of recalcitrant seeds of the tropical species, Avicennia marina. These are: 1) whether fungal infection reduces storage lifespan; 2) whether seeds become more susceptible to fungal attack during storage and whether they posses defence mechanisms that might suppress fungal proliferation in hydrated storage (production of antifungal compounds and 13-1,3-glucanase (EC 3.2.1.39) and chitinase (EC 3.2.1.14)] and 3) whether it is possible to discriminate ultrastructurally between inherent deteriorative changes and those that are fungally-induced. 1) The data indicate unequivocally that if fungal activity is curtailed, then the hydrated storage lifespan of A. marina seeds can be considerably extended. 2) When inoculated immediately with F. moniliforme, newly harvested seeds were extremely susceptible to the adverse effects of the fungus, while seeds that had been wet-stored for 4 days showed a considerably heightened resilience to the effects of the fungus prior to inoculation. The enhanced resilience, although declining, persisted in seeds stored hydrated for up to 10 days prior to inoculation, being lost after 12 days. This finding was supported by significant increase in 13-1,3-glucanase and chitinase and in antifungal compound production during 10 days of wet storage. After 14 days of wetstorage, seeds become more susceptible to the effects of fungusthanthose in the newly harvested condition. 3) The resilience of seeds that had been stored in the short-term was associated with ultrastructural changes indicative of enhanced metabolic activity associated with the onset of germination (e.g. increase in vacuolation, well-developed mitochondria and endomembrane system [ER and Golgi bodies]). However, with sustained stress associated with wet-storage IV conditions, the seeds became increasingly badly affected by the fungus, showing some ultrastructural fungally-induced abnormalities (e.g. nuclear lobing, presence of lipid bodies and prevalence of Golgi bodies that had many associated vesicles) and a decrease in 13-1,3-glucanase and chitinase activity. It is suggested that the decreased susceptibility of A. marina seeds during short-term storage relies on the ability to create an antifungal environment prior to infection (through synthesis and accumulation of pre-formed and induced antifungal compounds and antifungal enzymes), which would also be an effective strategy during germination in the natural environment. / Thesis (Ph.D.)-University of KwaZulu-Natal, Durban, 2004

Achieving long-term storage of germplasm is critical for the conservation of plant biodiversity. Seed storage practices require that degradative reactions causing ageing be limited. By reducing the water content, cytoplasmic viscosity is increased to levels that minimise deteriorative reactions. Reducing the storage temperature additionally increases the storage lifespan by further reducing the rate at which such deleterious processes occur. Two broad categories of seeds can be distinguished based on their storage behaviour. Orthodox seeds are desiccation-tolerant; generally shed in the dry state and are metabolically quiescent. Such seeds are usually stored at low water contents (e.g. 5%), and their high cytoplasmic viscosity prevents freezing damage during cooling to subzero temperatures. On the other hand, desiccation-sensitive (recalcitrant) seeds do not undergo a maturation-drying phase, they are metabolically active at shedding, and sensitive to extreme or prolonged drying. Accordingly, recalcitrant seeds cannot be stored under conventional conditions because they do not survive drying to low water contents and are damaged by sub-zero temperatures, even when dried to the lowest water content tolerated. Therefore, procedures that facilitate harmless drying and cooling to low temperatures are required to achieve long-term storage of recalcitrant germplasm. Recalcitrant seeds that are dried rapidly can attain relatively lower water contents without injury. However, these seeds are usually large and this limits the drying rates that can be achieved even under favourable conditions. Isolating embryonic axes from the rest of the seed facilitates faster drying, and a consequent reduction in the water content at which damage occurs. In axes of many species, the level of drying attained before lethal desiccation damage occurs is sufficient to limit freeZing damage during cryogenic exposure and facilitate survival in vitro. However, many others are damaged when dried to water contents that preclude freezing, and also are killed if cooled to sub-zero temperatures at higher water contents. In such instances, the window of permissible water contents leading to survival may be small or nonexistent. A basic premise explored in this thesis is that by restricting the growth of intracellular ice crystals using increasingly rapid cooling rates, the range of permissible water contents can be widened, facilitating survival of axes at higher water contents. An overview of the problems associated with the long-term storage of recalcitrant germplasm, and the rationale behind such rapid cooling approach are presented in Chapter 1 of the present thesis. Subsequent chapters report investigations on the effects of variables required to dry and cryopreserve embryonic axes with minimum damage, in keeping with this approach. Collectively, those studies aimed at establishing a robust cryopreservation procedure for the conservation of recalcitrant germplasm with broad applicability across species. The approach presently adopted entailed manipulating the water content of excised axes using rapid drying to discrete water content ranges, and also using different methods to cool axes to cryogenic temperatures at various rates. The calorimetric properties of water in axes were investigated for Camellia sinensis (L.) O. Kuntze using differential scanning calorimetry (DSC). For all species, the effect of any drying or cooling treatment tested was determined by assessing the survival of axes in vitro, which provided the most reliable indicator of cellular damage. Additionally, the effects of different treatments upon the structural and functional integrity of axes were assessed using light and electron microscopy as well as measurement of electrolyte leakage. The studies undertaken are presented in a similar sequence to that in which they took place during the course of the experimental phase of this work. These are summarised below. Partial drying plays a pivotal role in the approach developed, and microscopy has contributed towards increasing present understanding of desiccation damage. Microscopy was used to determine the effects of drying rate upon the ultrastructure of recalcitrant axes. It was necessary to find reliable protocols to prepare specimens for light and electron microscopy that did not alter the architecture of the cells in the dry state. Freeze-substitution and conventional aqueous fixation were compared in Chapter 2 using variously dried material from three species. The results obtained revealed that an unacceptably high extent of artefactual rehydration occurs during aqueous fixation, and highlight the need for anhydrous processing of dehydrated samples. Significantly, that study also revealed that many cellular events commonly associated with desiccation damage (e.g. withdrawal, tearing and/or vesiculation of the plasmalemma) are not seen in freeze-substituted preparations, and are likely artefacts of aqueous fixation. Freeze-substitution was used subsequently (Chapter 3) to assess the effects of slow drying (2 - 3 days) or rapid drying (min) upon the survival of embryonic axes of jackfruit (Artocarpus heterophyllus Lamk.) Results confirmed the beneficial effects of rapid drying, and also provided insights into ultrastructural changes and probable causes underlying cellular damage that occur during a drying/rehydration cycle. Efforts subsequently focused on determining the effect of cooling rate upon survival of recalcitrant axes at various water contents. The study on embryonic axes of recalcitrant camellia sinensis (tea; Chapter 4) tested the hypothesis that rapid cooling facilitates survival of axes at higher water content by restricting the growth of ice crystals to within harmless dimensions. The presence of sharp peaks in DSC melting thermograms was indicative of decreased survival in vitro. These peaks were attributed to the melting of ice crystals sufficiently large to be detected by DSC as well as to cause lethal damage to axes. Increasing the cooling rate from 10°C min-1 to that attained by forcibly plunging naked axes into sub-cooled nitrogen increased the upper limit of water content facilitating survival in vitro from c. 0.4 to 1.1 - 1.6 g H20 g-1 (dry mass [dmb]). Subsequent studies tested whether a similar trend occurred in other recalcitrant species cooled under similar conditions. In order to investigate further the relationship between water content, cooling rate and survival it was necessary to achieve cooling rates reproducibly, and to quantify these reliably. The plunging device required to achieve rapid cooling, and instruments required to measure the cooling rates attained, are described in Chapter 5. That study investigated the effects of cryogen type, depth of plunge and plunging velocity on the cooling rates measured by thermocouples either bare or within tissues of similar size and water content as encountered in cryopreservation experiments. This plunger was used in subsequent studies to achieve the fastest cooling conditions tested. Favourable cooling conditions were selected, and the efficacy of this procedure to cryopreserve relatively large axes was tested (Chapter 6) using embryonic axes of horse chestnut (Aesculus hippocastanum L.) Axes at water contents above c. 0.75 g g-1 could not be cooled faster than c. 60°C S-1, but cooling rates of axes below this water content increased markedly with isopentane, and to a lesser extent with subcooled nitrogen. Axes were killed when cooled at water contents above 1.0 g g-1 but survived fully (albeit abnormally) when cooled in isopentane between 1.0 and 0.75 g g-1. Complete survival and increasingly normal development was attained at water contents below 0.75 g g-1, especially if isopentane was used. The study on horse chestnut axes emphasised that water content and cooling rate are co-dependent during non-equilibrium cooling. Accordingly, that study could not determine whether survival at lower water contents increased because of the corresponding increase in cooling rates measured, or because of the higher cytoplasmic viscosity that resulted from drying. That uncertainty was addressed by the study discussed in Chapter 7, using axes of the trifoliate orange (Poncirus trifoliata [L.] RAF.) That study investigated the effect of cytoplasmic viscosity upon survival of axes cooled and warmed at different rates. Survival and normal development was high at lower water contents, and seemingly independent of cooling rate at about 0.26 g g-1. At higher water contents the range of cooling rates facilitating survival became narrower and displaced towards higher cooling rates. This study revealed two conspicuous inconsistencies that questioned the beneficial effect of rapid cooling. Firstly, the fastest cooling rates did not necessarily facilitate the highest survival. Secondly, survival of fully hydrated axes was higher when cooled under conditions that encouraged - rather than restricted - the growth of intracellular ice crystals. These inconsistencies were explored further using embryonic axes of silver maple (Acer saccharinum L.). Freeze-fracture replicas and freeze-substitution techniques provided valuable insights into the way in which ice crystals were distributed in cells cooled using different methods at rates ranging between 3.3 and 97°C S-1. Extensive intracellular freezing was common to all treatments. Unexpectedly, fully hydrated axes not only survived cryogenic exposure, but many axes developed normally when cooled using the relatively slower methods (77 and 3.3°C S-1) if warming was rapid. The most conspicuous ultrastructural difference between plunge cooling and the relatively slower methods was the exclusion of ice from many intracellular compartments in the latter. It is possible that even the fastest warming cannot prevent serious cellular damage if ice crystals form within such 'critical' compartments. It is proposed that the intracellular location of ice is a stronger determinant of survival that the size attained by ice crystals. The study of A. saccharinum also investigated the recovery of axes cooled fully hydrated either rapidly (97°C S-1) or slowly (3.3°C S-1). This facet of the study showed that cell lysis became apparent immediately after warming only where damage was most extensive. In other cells damage became apparent only after 2.5 to 6 h had elapsed, thus cautioning against inferring survival from the ultrastructural appearance of cells immediately after warming. Microscopy enabled cell repair as well as the pattern of growth of cryopreserved tissues to be appraised at the cellular, tissue and organ levels. Similar studies are required to understand further the nature of freezing damage, and how those events affect cell function. The salient trends observed in previous chapters are brought together in Chapter 9. / Thesis (Ph.D.)-University of Natal, Durban, 2002.