The encroachment of rangelands by woody plants causes an imbalance in the grass:bush ratio leading to decline in ecosystem services including grazing capacity, biodiversity, and water yield. This study assessed the historical changes in woody cover, and the effects of bush clearing on water loss, atmospheric carbon uptake, herbaceous vegetation, and soil seed bank (SSB) dynamics. The study was conducted at Makapanstad (Radi and Maseding) and Kgomokgomo rangelands, North-West Province, South Africa. Landsat imagery was used to assess woody cover over 34 years (1984 - 2018) in a total area of 16 397 ha. Evapotranspiration (ET) and gross primary productivity (GPP) were assessed in cleared sites and their adjacent uncleared sites from 2013 to 2018. Woody plant densities, herbaceous composition, ground cover and biomass production were assessed in 24, 5×5-m plots distributed equally in three 2500-m2 bush-encroached blocks in each rangeland in February 2016. Three samples for soil nutrient analysis were collected 20-cm deep in each plot. Thereafter, half (1250-m2) of each block was mechanically cleared to make three replicates of cleared and uncleared microsites. Post-treatment survey of vegetation was conducted in February 2017. SSB was assessed for samples collected in April 2016 before bush clearing and in August, October and December 2016, and April and August 2017 in paired microsites. In 1984, herbaceous cover (humid + arid grasslands) dominated, accounting for 36% of the total area followed by woody cover (shrublands + bushlands) and unvegetated cover (bare soils + abandoned croplands), with respective proportions of 33 and 31%. There were substantial land cover changes over time, with shrub cover increasing linearly (r2 = 0.94, p < 0.05) at a rate of 0.26% year-1, increasing the total woody cover (TWC) to 38% in 2018. Cropland abandonment was the main driver of increase in shrub cover in the study area. Herbaceous cover declined with increase in TWC (r2 = 0.69), whereas bare soil cover increased (r2 = 0.70) with TWC over time. The main encroachers were Vachellia tenuispina at Radi, Vachellia tortilis at Maseding and Dichrostachys cinerea and V. tortilis at Kgomokgomo. Most of the woody species correlated with total N and clay content (r = 0.96; p < 0.05) in black vertic clay soils of Radi and Maseding, whereas non-leguminous species correlated with total N and C:N in the red-yellow apedal soils of Kgomokgomo. ET and GPP varied interannually in all rangelands and both declined significantly (p < 0.05) after bush clearing at Radi and Kgomokgomo but not at Maseding. ET increased with GPP in cleared (r2 = 0.50-0.59) and uncleared sites (r2 = 0.82-0.93) at Radi and they showed a strong relationship (r2 >0.70) in both sites at Maseding and Kgomokgomo. ET ranged from 0.26-0.46 and 0.48-0.97 Kg H2O m-2 day-1 in cleared and uncleared sites, respectively, at Radi and from 0.44-0.98 and 0.63-1.24 Kg H2O m-2 day-1 at Kgomokgomo. Total SSB densities (constituted mostly by early successional species) increased significantly (p < 0.05) to 2470, 1872 and 693 seeds m-2 at Radi, Maseding and Kgomokgomo, respectively, in cleared microsites in August 2017. Similarly, biomass production (BP) and basal cover (BC) were highest in cleared relative to uncleared microsites. Overall, increase in woody cover reduced herbaceous cover and this calls for bush control in the studied rangelands. The decline in ET after bush clearing suggested that bush clearing could be used to increase rangeland water yield. Increased SSB densities, BP and BC indicated that bush clearing facilitates passive restoration. / Thesis (PhD (Pasture Science))--University of Pretoria, 2020. / Agricultural Research Council / National Research Foundation / Plant Production and Soil Science / PhD (Pasture Science) / Unrestricted

Understanding how organisms respond to climatic variability and novel conditions is becoming an increasingly important task for ecologists. For ectotherms in the northern hemisphere, the response to cold is of special interest, considering that poleward range expansion events and increasing variability of temperatures during winter are already being observed as consequences of a warming planet. Though direction of change in physiological variables in response to cold is well studied in ectotherms, the extent to which traits can change and the rate at which they can change is not. We compared the extent and rate of change in cold tolerance (CTmin) between two long-term captive populations of the Italian wall lizard (Podarcis siculus) during a lab cold-acclimation treatment. Heat tolerance (CTmax), thermal preference (Tpref), temperature dependent rates of oxygen consumption (SMRO2), and temperature dependent rates of water loss (EWL) were also compared between Italian wall lizards previously introduced to Long Island, NY and San Pedro, CA before and after the lab cold acclimation treatment. Because our study coincided with a cold snap during the spring 2018 season for the San Pedro, CA population, we also studied the effects of cold acclimatization on wild lizards from the CA population. After initial lab acclimation of the lizards to laboratory conditions, SMRO2 at 15°C and EWL at 10°C were higher in NY lizards compared to CA lizards. Lizards from the two populations did not differ in any other variables measured before the cold acclimation treatment. We found that lizards from the NY population experienced an 80% decrease in CTmin following a switch from 20°C:18°C to 17.5°C:16°C (12h light:12h dark) acclimation treatment. Lizards from the CA population did not decrease CTmin in response to the same cold acclimation treatment. Overall, NY lizards decreased CTmin, CTmax, and Tpref following cold acclimation, whereas CA lizards decreased CTmax only. Wild CA lizards decreased CTmax following the cold spring 2018 season in a manner similar to that of lab acclimated NY and CA lizards, suggesting that these lizards do not maintain a high CTmax when the environment is unlikely to expose them to high temperatures. Thermal sensitivity (Q10) of SMRO2 and EWL was lower in NY lizards, suggesting physiological adaptation to fluctuation in diurnal temperatures. The ratio of CO2 produced to O2 consumed (respiratory exchange ratio, RER) measured at 15°C increased in NY lizards following cold acclimation suggesting an increased use of carbohydrates and/or an increased production of lipids in the colder conditions. These responses in combination with the higher observed plasticity in NY lizards are in accordance with the climatic variability hypothesis, which predicts that organisms from more variable climates will be better adapted to physiologically respond to variable conditions. The higher capacity for physiological plasticity may explain the relatively high success of P. siculus in NY and other northern U.S. states. By describing the rate of change of CTmin during cold acclimation we hope to better understand how these lizards minimize the risk of low temperature exposure during winter. We ultimately hope to incorporate the rate at which cold tolerance can change into predictions of species distributions and hypothesis tests investigating the relationship between climatic variability and the rate at which animals can exhibit plasticity.

Chapter 1: Animals can respond to extreme climate by behaviorally avoiding it, or by physiologically coping with it. We understand behavioral thermoregulation and physiological thermal tolerances, but water balance has largely been neglected. Climate change includes both global warming and changes in precipitation regimes, so improving our understanding of organismal water balance is increasingly urgent. We assessed the hydric physiology of endangered Blunt-nosed Leopard Lizards (Gambelia sila) by measuring cutaneous evaporative water loss (CEWL), plasma osmolality, body mass, and body condition throughout their active season. On average, G. sila had low CEWL that is likely desert-adaptive, and high plasma osmolality that is indicative of dehydration. Given that our study was in a drought year, it is reasonable to believe that every lizard measured was dehydrated to a degree. We hypothesized that throughout the G. sila active season, as their habitat got hotter and drier, G. sila would become increasingly dehydrated and watertight. Instead, CEWL and plasma osmolality showed minimal change for females and nonlinear change for males, which we hypothesize is connected to sex-specific reproductive behaviors and changes in food availability. We also measured thermoregulation and microhabitat use, expecting that more hydrated lizards would have higher body temperature, better thermoregulatory accuracy, and spend more time aboveground. However, we found no effect of CEWL, plasma osmolality, body mass, or body condition on these thermal and behavioral metrics. We posit either that G. sila tolerate dehydration to maintain activity during their brief active season, or that because every lizard was dehydrated due to the drought, they all experienced equally constrained thermoregulation and microhabitat use. Finally, G. sila spend considerable time underground in burrows, and we believe burrows serve as essential hydric, not only thermal, refugia. Our findings suggest that these lizards might benefit from artificial humid refugia and supplemental hydration, especially during drought. Chapter 2: Testing acclimation plasticity informs our understanding of functional biodiversity and applies to conservation management amidst our rapidly changing climate. While there is a wealth of research on the plasticity of thermal and hydric physiology in response to temperature acclimation, there is a comparative gap for research on acclimation to different hydric regimes, as well as the interaction between water and temperature. We sought to fill this gap by acclimating Western Fence Lizards (Sceloporus occidentalis) to experimental climate conditions (crossed design of Hot or Cool, Dry or Humid) for eight days, and measuring cutaneous evaporative water loss (CEWL), plasma osmolality, hematocrit, and body condition before and after acclimation under common conditions. CEWL changed plastically in response to the different climates, with lizards acclimated to Hot Humid conditions experiencing the greatest increase in CEWL. Change in CEWL among individuals was negatively related to treatment vapor pressure deficit. Plasma osmolality, hematocrit, and body condition all showed greater changes in response to temperature than to humidity or vapor pressure deficit. CEWL and plasma osmolality were positively related across treatment groups before acclimation and within treatment groups after acclimation, but the two variables showed different responses to acclimation, suggesting that they are interrelated but governed by different mechanisms. This study is among just a small number of studies that assess more than one metric of hydric physiology and that test the interactive effects of temperature and humidity. Such measurements will be essential for predictive models of activity and survival for animals under climate change.

Yes / Soft silicone's flexibility, adhesive capacity and non-toxic, non-odourous and hypoallergenic nature have made it an established material for adhesive and protective therapeutic devices. In wound care, silicone is a component of contact layer dressings for superficial wounds and silicone gel sheeting for reducing the risk of scarring, as well as of barriers for incontinence-associated dermatitis. Regarding stoma accessories, silicone is established in barrier films to prevent contact dermatitis, adhesive removers to prevent skin stripping and filler gels to prevent appliance leaks. Until recently, silicone has not been used in stoma appliances flanges, as its hydrophobic nature has not allowed for moisture management to permit trans-epidermal water loss and prevent maceration. Traditional hydrocolloid appliances manage moisture by absorbing water, but this can lead to saturation and moisture-associated skin damage (MASD), as well as increased adhesion and resultant skin tears on removal, known as medical adhesive-related skin injury (MARSI). However, novel silicone compounds have been developed with a distinct evaporation-based mechanism of moisture management. This uses colloidal separation to allow the passage of water vapour at a rate equivalent to normal trans-epidermal water loss. It has been shown to minimise MASD, increase wear time and permit atraumatic removal without the use of adhesive solvents. Trio Healthcare has introduced this technology with a range of silicone-based flange extenders and is working with the University of Bradford Centre for Skin Sciences on prototype silicone-based stoma appliance flanges designed to significantly reduce the incidence of peristomal skin complications, such as MARSI and MASD. It is hoped that this will also increase appliance wear time, reduce costs and improve patient quality of life.

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Skin is the largest immune organ and an ideal site to administer vaccines. However, by nature, skin is not permeable to antigens, which are macromolecules. The major hurdle in skin permeation is the outermost stratum corneum layer. Microneedles have proven feasible to create micron-sized channels in the epidermis of the skin, through which protein and plasmid DNA antigens can penetrate into the viable skin epidermis and dermis. However, the immune responses induced by microneedle-mediated transcutaneous immunization with protein or plasmid DNA alone are generally weak, and a vaccine adjuvant is often required to induce strong immune responses. Data from numerous previous studies have shown that nanoparticles as a vaccine carrier can significantly enhance the immunogenicity of antigens, but the feasibility of utilizing nanoparticles as a vaccine carrier to enhance the immune responses induced by microneedle-mediated transcutaneous immunization has rarely been studied. In this dissertation, using protein antigen (OVA) chemically conjugated onto the surface of solid-lipid nanoparticles and plasmid DNA (pCMV-beta, pVax/opt-BoNT/C-Hc50, and pCI-neo-sOVA) physically coated on the surface of cationic polymeric nanoparticles, we showed that the immune responses induced by microneedle-mediated transcutaneous immunization with protein antigens or plasmid DNA vaccines are significantly enhanced by delivering the proteins and plasmid DNA with nanoparticles. Importantly, microneedle-mediated transcutaneous immunization with proteins or plasmid DNA induces not only systemic immune responses, but also mucosal immune responses. In addition, it is generally believed that microneedles are safe. However, it remained unclear whether the micropores created by microneedles on the skin will also facilitate the permeation of microbes such as bacteria into the skin. In this dissertation, we also designed an unique ex vivo model to evaluate the permeation of live bacteria through mouse skin pretreated with microneedles. The results demonstrated that the risk of potential bacterial infection associated with microneedle treatment is not greater than that associated with a hypodermic needle injection. / text

Moisturizers are used in various types of dry skin disorders, but also by people with healthy skin. It is not unusual that use of moisturizers is continued for weeks, months, or even years. A number of moisturizers have been shown to improve the skin barrier function, while others to deteriorate it, but the reason for observed effects remains unknown. Further understanding of the mechanism by which long-term treatment with moisturizers influences the skin barrier would have clinical implications, as barrier-deteriorating creams may enhance penetration of allergens or irritants and predispose to dry skin and eczema, while barrier-improving ones could reduce many problems. The present research combined non-invasive techniques with analyses of skin biopsies, allowing studies of the epidermis at molecular and cellular level. Test moisturizers were examined on healthy human volunteers for their effect on the skin barrier, with regard to such factors as pH, lipid type, and presence of a humectant, as well as complexity of the product. After a 7-week treatment with the moisturizers, changes in transepidermal water loss, skin capacitance, and susceptibility to an irritant indicated a modified skin barrier function. Moreover, the mRNA expression of several genes involved in the assembly, differentiation and desquamation of the stratum corneum, as well as lipid metabolism, was altered in the skin treated with one of the moisturizers, while the other moisturizer induced fewer changes. In conclusion, long-term use of moisturizers may strengthen the barrier function of the skin, but also deteriorate it and induce skin dryness. Moisturizers have also a significant impact on the skin biochemistry, detectable at molecular level. Since the type of influence is determined by the composition of a moisturizer, more careful selection of ingredients could help to design moisturizers generating a desired clinical effect, and to avoid ingredients with a negative impact on the skin.

Atopic dermatitis (AD), ichthyosis vulgaris (IV), and X-linked recessive ichthyosis (XLRI) are characterized by dry skin and impaired skin barrier. AD and IV are related to loss-of-function mutations in FLG (encoding filaggrin), whereas XLRI is caused by deletions or inactivating mutations in the steroid sulphatase gene (STS). Patients regularly use moisturizing creams, but little is known about the creams’ effects on the skin barrier. The present work combines objective scorings, non-invasive techniques, and molecular analyses of skin biopsies to characterize the skin in 57 patients with AD, IV, or XLRI, and in 14 healthy controls. Patients were classified according to their FLG and STS mutation status: AD with FLG+/+ (n = 14), AD with FLG+/– (n = 14), AD/IV with FLG–/– (n = 15), and XLRI with STS– (n = 14), as well as one man with a novel point mutation. Assessments were conducted at baseline and after four weeks of treatment with three different moisturizers applied to volar forearm skin. At baseline, dryness scoring and non-invasive assessments verified impaired skin barrier function in all patients. In patients with AD/IV, microarray analysis identified 300–3000 up- or downregulated mRNA transcripts involved in signalling pathways important for inflammation and barrier repair. The skin phenotype and number of altered transcripts were correlated with the FLG mutation status, with FLG–/– patients displaying the highest transepidermal water loss (TEWL) and the most altered transcript levels. In contrast, despite an equally dysfunctional skin barrier, only limited changes in mRNA transcripts occurred in XLRI patients. Treatment with moisturizers improved skin dryness similarly in all groups, but TEWL behaved differently: it decreased slightly in the AD/IV group and increased in the XLRI group, especially after urea treatment. Only minute effects on skin pH and mRNA expression were observed. In conclusion, FLG mutations elicit pro-inflammatory mechanisms probably aimed at restoring barrier competence. This does not occur in patients with XLRI, presumably because STS deficiency automatically increases the barrier thickness. Moisturizing treatment improves skin dryness in patients with AD, IV, or XLRI, but does not seem to normalize the altered epidermal gene expression profile in AD/IV patients.