Small cationic DDA:TDB liposomes as protein vaccine adjuvants obviate the need for TLR agonists in inducing cellular and humoral responses

Most subunit vaccines require adjuvants in order to induce protective immune responses to the targeted pathogen. However, many of the potent immunogenic adjuvants display unacceptable local or systemic reactogenicity. Liposomes are spherical vesicles consisting of single (unilamellar) or multiple (multilamellar) phospholipid bi-layers. The lipid membranes are interleaved with an aqueous buffer, which can be utilised to deliver hydrophilic vaccine components, such as protein antigens or ligands for immune receptors. Liposomes, in particular cationic DDA:TDB vesicles, have been shown in animal models to induce strong humoral responses to the associated antigen without increased reactogenicity, and are currently being tested in Phase I human clinical trials. We explored several modifications of DDA:TDB liposomes--including size, antigen association and addition of TLR agonists--to assess their immunogenic capacity as vaccine adjuvants, using Ovalbumin (OVA) protein as a model protein vaccine. Following triple homologous immunisation, small unilamellar vesicles (SUVs) with no TLR agonists showed a significantly higher capacity for inducing spleen CD8 IFN? responses against OVA in comparison with the larger multilamellar vesicles (MLVs). Antigen-specific antibody reponses were also higher with SUVs. Addition of the TLR3 and TLR9 agonists significantly increased the adjuvanting capacity of MLVs and OVA-encapsulating dehydration-rehydration vesicles (DRVs), but not of SUVs. Our findings lend further support to the use of liposomes as protein vaccine adjuvants. Importantly, the ability of DDA:TDB SUVs to induce potent CD8 T cell responses without the need for adding immunostimulators would avoid the potential safety risks associated with the clinical use of TLR agonists in vaccines adjuvanted with liposomes.

Functionalisation of ethylene-propylene rubber with glycidyl methacrylate in the presence of comonomer and in situ compatibilisation of PET/f-EPR blends

The main aim of this work was two fold, firstly to investigate the effect of a highly reactive comonomer, divinylbenzene (DVB), on the extent of melt grafting of glycidyl methacrylate (GMA) on ethylene-propylene rubber (EPR) using 2,5-dimethyl-2,5-bis-(tert-butyl peroxy) hexane (Trigon ox 101, Tl 01) as a free radical initiator, and to compare the results with a conventional grafting of the same monomer on EPR. To achieve this, the effect of processing conditions and chemical composition including the concentration of peroxide, GMA and DVB on the extent of grafting was investigated. The presence of the comonomer (DVB) in the grafting process resulted in a significant increase in the extent of the grafting using only a small concentration of peroxide. It was also found that the extent of grafting increased drastically with increasing the DVB concentration. Interestingly, in the comonomer system, the extent of the undesired side reaction, normally the homopolymerisation of GMA (polyGMA) was shown to have reduced tremendously and in most cases the level of polyGMA was immeasurable in the samples. Compared to a conventional EPR-g-GMACONV (in the absence of a comonomer), the presence of the comonomer DVB in the grafting system was shown to result in more branching and crosslinking (shown from an increase in melt flow index (MFI) and torque values) and this was paralleled by an increase in DVB concentration. In contrast, the extent of grafting in conventional system increased with increasing the peroxide concentration but the level of grafting was much lower than in the case of DVB. Homopolymerisation of GMA and excessive crosslinking of EPR became dominant at high peroxide concentration and this. reflects that the side reactions were favorable in the conventional grafting system. The second aim was to examine the effect of the in-situ functionalised EPR when used as a compatibiliser for binary blends. It was found that blending PET with functionalised EPR (ƒ-EPR) gave a significant improvement in terms of blend morphology as well as mechanical properties. The results showed clearly that, blending PET with ƒ-EPRDVB (prepared with DVB) was much more effective compared to the corresponding PET/ƒ-EPRCONV (without DVB) blends in which ƒ-EPRDVB having optimum grafting level of 2.1 wt% gave the most pronounced effect on the morphology and mechanical properties. On the other hand, blends of PET/ƒ-EPRDVB containing high GMA/DVB ratio was found to be unfavorable hence exhibited lower tensile properties and showed unfavorable morphology. The presence of high polyGMA concentration in ƒ-EPRCONV was found to create damaging effect on its morphology, hence resulting in reduced tensile properties (e.g. low elongation at break). However, the use of commercial terpolymers based on ethylene-methacrylate-glycidyl methacrylate (EM-GMA)or a copolymer of ethylene-glycidyl methacrylate (E-GMA) containing various GMA levels as compatibilisers in PET/EPR blends was found to be more efficient compared to PET/EPR/ƒ-EPR blends with the former blends showing finer morphology and high elongation at break. The high efficiency of the terpolymers or copolymers in compatibilising the PET/EPR blends is suggested to be partly, higher GMA content compared to the amount in ƒ-EPR and due to its low viscosity.

High plasticity of loggerhead nesting site fidelity: from using repeatedly the same small beach during different seasons to deposit consecutive nests in different islands distant more than 100 km

[EN] Sea turtles exhibit a strong natal homing associated with a high nesting site philopatry. Mark-recapture and genetic studies confirm these patterns suggesting differences among and within species. In the present study, we have analysed the degree of nest site fidelity of a loggerhead population nesting on the islands of Cape Verde and have evaluated the existence of intrapopulational variability in this trait. The loggerhead is the only sea turtle species that nests in Cape Verde, and 15,000–25,000 nests per season have been estimated for the whole archipelago.

Evidence from genetic and Lagrangian drifter data for transatlantic transport of small juvenile green turtles

[EN] Aim: A key life-history component for many animals is the need for movement between different geographical locations at particular times. Green turtle (Chelonia mydas) hatchlings disperse from their natal location to spend an early pelagic stage in the ocean, followed by a neritic stage where small juveniles settle in coastal areas. In this study, we combined genetic and Lagrangian drifter data to investigate the connectivity between natal and foraging locations. In particular, we focus on the evidence for transatlantic transport.

Opportunities and constraints of transhumant small ruminant production in the Chinese Altay Mountains

This PhD thesis focuses on current livelihoods of agro-pastoral livestock keepers, their animal nutrition, herd and rangeland management strategies. It thereby aims to contribute to sustainable rangeland management, livestock production and household income in Qinghe county of the Chinese Altay Mountain region, located in Xinjiang Uyghur Autonomous Region, PR China. In its first part the study characterizes the socio-economic situation and agricultural practices of agro-pastoralists through structured household interviews. The second part provides insights into the grazing behaviour and feed intake of small ruminants on seasonal pastures in this region, and into the quantitative and qualitative biomass offer on natural rangelands. The third part analyses the reproductive performance and annual growth of the local sheep and goat herds, and, by modelling improved feeding and culling strategies, tests herd management options that potentially improve the monetary output per female herd animal without increasing the pressure onto natural rangelands. Taken together, the results of the study suggest that, despite an increase and intensification of cropping and vegetable gardening in the region of Qinghe, livestock rearing is still the major livelihood strategy both in terms of prevalence and relative importance. However, livestock keeping is challenged by low biomass production on rangelands, due to the combined impact of high climate variability and highly localized grazing pressure on the seasonal pastures. Though government regulations try to tackle the latter aspect, their implementation is sometimes difficult. Alternatives to strict regulation of grazing periods and animal numbers on seasonal pastures are, in the case of goats, more rigorous culling strategies and, in the case of sheep and goats, strategic supplementation of the animals in the winter and spring season. However, for the latter strategy to become economically viable, an improvement of live animal and meat marketing options and an investment in local meat processing facilities that add value to the carcasses is needed. As the regional cities grow rapidly, the potential market to absorb diverse and good quality meat products is there, along with the road network connecting Qinghe county to the regional capital. Such governmental measures will not only create new job opportunities in the region but also benefit the cash income of pastoralists in this westernmost region of China.

Evidence from genetic and Lagrangian drifter data for transatlantic transport of small juvenile green turtles

[EN] Green turtle hatchlings disperse away from their natal location to spend an early pelagic stage in the ocean, followed by a neritic stage where small juveniles settle in coastal areas. Here, we combined genetic and Lagrangian drifter data to investigate the connectivity between natal and foraging locations; particularly focussing on the evidence for transatlantic transport. Our results supported the general hypothesis that turtles tend to select foraging areas ‘closest-to-home’.

GPS - PET : aplicación web para la geolocalización de mascotas

[ES]Este proyecto trata de desarrollar una aplicación web en Ruby on Rails, donde el usuario pueda visualizar en “tiempo real” la ubicación de su animal haciendo uso de su identificador con GPS (microchip con GPS), si existiera tal dispositivo, o un dispositivo de geolocalización GPS incrustado en el collar del perro, para hacer un seguimiento del mismo cuando otra persona que ha sido contratada lo pasea. Al no disponer de microchips o collares con GPS, así como los respectivos animales para hacer el estudio y, por tanto, su posterior seguimiento, se realizará una simulación del mismo, de ahí que el tiempo real esté puesto entre comillas.

Novel Anti-Campylobacter Compounds Identified Using High Throughput Screening of a Pre-selected Enriched Small Molecules Library

Campylobacter is a leading cause of foodborne bacterial gastroenteritis worldwide and infections can be fatal. The emergence of antibiotic-resistant Campylobacter spp. necessitates the development of new antimicrobials. We identified novel anti-Campylobacter small molecule inhibitors using a high throughput growth inhibition assay. To expedite screening, we made use of a “bioactive” library of 4,182 compounds that we have previously shown to be active against diverse microbes. Screening for growth inhibition of Campylobacter jejuni, identified 781 compounds that were either bactericidal or bacteriostatic at a concentration of 200 µM. Seventy nine of the bactericidal compounds were prioritized for secondary screening based on their physico-chemical properties. Based on the minimum inhibitory concentration against a diverse range of C. jejuni and a lack of effect on gut microbes, we selected 12 compounds. No resistance was observed to any of these 12 lead compounds when C. jejuni was cultured with lethal or sub-lethal concentrations suggesting that C. jejuni is less likely to develop resistance to these compounds. Top 12 compounds also possessed low cytotoxicity to human intestinal epithelial cells (Caco-2 cells) and no hemolytic activity against sheep red blood cells. Next, these 12 compounds were evaluated for ability to clear C. jejuni in vitro. A total of 10 compounds had an anti-C. jejuni effect in Caco-2 cells with some effective even at 25 µM concentrations. These novel 12 compounds belong to five established antimicrobial chemical classes; piperazines, aryl amines, piperidines, sulfonamide and pyridazinone. Exploitation of analogues of these chemical classes may provide Campylobacter specific drugs that can be applied in both human and animal medicine.

Animal Models of Friedreich Ataxia

Friedreich ataxia (FRDA) is the most common form of autosomal-recessive ataxia. Common nonmotor features include cardiomyopathy and diabetes mellitus. At present, no effective treatments are available to prevent disease progression. Age of onset varies from infancy to adulthood. In the majority of patients, FRDA is caused by intronic GAA expansions in FXN, which encodes a highly-conserved small mitochondrial matrix protein, frataxin. A mouse model of FRDA has been difficult to generate because complete loss of frataxin causes early embryonic lethality. Although there are some controversies about the function of frataxin, recent biochemical and structural studies have confirmed that it is a component of the multiprotein complex that assembles iron-sulfur clusters in the mitochondrial matrix. The main consequences of frataxin deficiency are energy deficit, altered iron metabolism, and oxidative damage.

A PRELIMINARY SURVEY OF ANIMAL HANDLING AND CULTURAL SLAUGHTER PRACTICES AMONG KENYAN COMMUNITIES: POTENTIAL INFLUENCE ON MEAT QUALITY

Kenya is composed of over 40 ethnic communities who practice varied methods of animal handling and slaughter. Socio-cultural and religious traditions have the potential to influence animal handling and slaughter practices. These influences have, however, not been documented in the literature as far as the author is aware. Also, the literature has documented the connection between the manner of animal treatment and meat quality, but this is rarely discussed in the literature in Kenya; this connection is important as it informs modern meat trade practices by Kenyans as they trade in the global arena. This survey aimed to mainly establish and document the animal slaughter practices among Kenyan communities, and, to also highlight any current provisions related to meeting modern animal welfare requirements, animal handling procedures in the meat trade and discuss their potential influence on meat quality available in commerce in Kenya. This preliminary study surveyed the slaughter practices among 10 different Kenyan communities through a semi-structured questionnaire, focus group discussions and individual interviews. The survey demonstrated that different Kenyan communities practice varied methods of animal slaughter depending on whether the animal being slaughtered is for public feasting, domestic consumption or commercial merchandizing. The Kenyan communities surveyed in this study depend mainly on males to slaughter livestock for females preparing it for domestic use using a number of instruments and methods. For small stock for domestic consumption, females may slaughter the animal except for Muslims whose males have to slaughter the animal with a special knife (a Khalef) according to Muslim rites to render it Halal. Large stock is invariably slaughtered by males irrespective of the community, and the manner of use of the carcass. Gender, age, religion, community and the size of the animal were the major determinants of the method of animal slaughter. The animal welfare issues highlighted in the survey and related to the handling and slaughter of livestock have important implications for meat quality during commercial merchandizing. There is an apparent need to provide education to herders, livestock handlers, employees and management in the livestock industry in Kenya on the relationship between animal welfare requirements, animal handling procedures and meat quality. Such awareness can potentially improve the quality and economic value of the meat available in commerce.