Physicochemical and molecular characterization of bacteriophages ΦSP-1 and ΦSP-3, specific for pathogenic Salmonella and evaluation of their potential as biocontrol agent

This thesis entitled Physicochemical and molecular characterization of bacteriophages ΦSP-1and ΦSP-3, specific for pathogenic Salmonella and evaluation of their potential as biocontrol agent . Salmonella were screened using standard methodologies from various environmental samples including chicken caecum. Salmonella strains, which were previously isolated and stocked in the lab, were also included in this study as host, for screening Salmonella specific lytic phages. The Salmonella strain in this study designated as S49 which helped in phage propagation by acting as host bacteria was identified as Salmonella enterica subsp. enterica by 16S rRNA gene analysis and serotyping . A total of three Salmonella specific phage named as ΦSP-1, ΦSP-2 and ΦSP-3 were isolated from chicken intestine samples via an enrichment protocol employing the double agar overlay method. ΦSP-1 and ΦSP-3 showing consistent lytic nature were selected for further study and were purified by repeated plating after picking of single isolated plaques from the lawns of Salmonella S49 plates. Both the phages produced small, clear plaques indicating their lytic nature. ΦSP-1 and ΦSP-3 were concentrated employing PEG-NaCl precipitation method before further characterization. The focus of present study was to isolate, characterize and verify the efficacy of lytic bacteriophages against the robust pathogen Salmonella, capable of surviving under various hostile conditions. Two phages, ΦSP-1 and ΦSP-3, belonging to two families, Podovoridae and Siphoviridae were isolated.

“Development of lymphoid cell culture system from Penaeus monodon and molecular approaches for its transformation

Unveiling the molecular and regulatory mechanisms that prevent in vitro transformation in shrimp remains elusive in the development of continuous cell lines, with an arduous history of over 25 years (Jayesh et al., 2012). Despite presenting challenges to researchers in developing a cell line, the billion dollar aquaculture industry is under viral threat. In addition, the regulatory mechanisms that prevent in vitro transformation and carcinoma in shrimps might provide new leads for the development of anti-ageing and anti-cancer interventions in human (Vogt, 2011) and in higher vertebrates. This highlights the importance of developing shrimp cell lines, to bring out effective prophylactics against shrimp viruses and for understanding the mechanism that induce cancer and ageing in human.. Advances in molecular biology and various gene transfer technologies for immortalization of cells have resulted in the development of hundreds of cell lines from insects and mammals, but yet not a single cell line has been developed from shrimp and other marine invertebrates. With this backdrop, the research described in this thesis attempted to develop molecular tools for induced in vitro transformation in lymphoid cells from Penaeus monodon and for the development of continuous cell lines using conventional and novel technologies to address the problems at cellular and molecular level.

Detection of sandal spike phytoplasma using immunological and molecular techniques

Spike disease in sandal is generally diagnosed by the manifestation of external symptoms. Attempts have been made to detect the diseased plants by determining the length/breadth ratio of leaves (lyengar, 1961) and histochemical tests using Mann's stain (Parthasarathi et al., 1966), Dienes' stain (Ananthapadmanabha et a/., 1973) aniline blue and Hoechst 33258 (Ghosh et a/., 1985, Rangaswamy, 1995). But most of these techniques are insensitive, indirect detection methods leading to misinterpretation of results. Moreover, to identify disease resistant sandal trees, highly sensitive techniques are needed to detect the presence of the pathogen. In sandal forests, several host plants of sandal like Zizyphus oenop/ea (Fig. 1.3) also exhibit the yellows type disease symptoms. Immunological and molecular assays have to be developed to confirm the presence of sandal spike phytoplasma in such hosts. The major objectives of the present work includes:In situ detection of sandal spike phytoplasma by epifluorescence microscopy and scanning electron microscopy.,Purification of sandal spike phytoplasma and production of polyclonal antibodies.,Amino acid and total protein estimation of sandal spike phytoplasma.,Immunological detection of sandal spike phytoplasma., Molecular detection of sandal spike phytoplasma.,Screening for phytoplasma in host plants of spike disease affected sandal using immunological and molecular techniques.

A facile and single pot strategy for the synthesis of novel naphthyridine derivatives under microwave irradiation conditions using ZnCl2 as catalyst, evaluation of AChE inhibitory activity, and molecular modeling studies

A series of novel naphthyridine derivatives 3 and 4 was prepared from substituted pyridine 2 and ketones using ZnCl2 as catalyst under microwave irradiation conditions. All the compounds were evaluated for AChE inhibitory activity and promising compounds 3d, 3e, 4b, and 4g was identified. Representative compounds 3d and 3e were found to show insignificant THLE-2 liver cell viability/toxicity. The binding mode between X-ray crystal structure of human AChE and compounds was studied using molecular docking method and fitness scores were found to be in good correlation with the activity data.

Differential induction, isolation, physicochemical and molecular characterization of temperate phages of environmental Vibrio cholerae as evidence of phage mediated Horizontal Gene Transfer

The resurgence of the enteric pathogen Vibrio cholerae, the causative organism of epidemic cholera, remains a major health problem in many developing countries like India. The southern Indian state of Kerala is endemic to cholera. The outbreaks of cholera follow a seasonal pattern in regions of endemicity. Marine aquaculture settings and mangrove environments of Kerala serve as reservoirs for V. cholerae. The non-O1/non-O139 environmental isolates of V. cholerae with incomplete ‘virulence casette’ are to be dealt with caution as they constitute a major reservoir of diverse virulence genes in the marine environment and play a crucial role in pathogenicity and horizontal gene transfer. The genes coding cholera toxin are borne on, and can be infectiously transmitted by CTXΦ, a filamentous lysogenic vibriophages. Temperate phages can provide crucial virulence and fitness factors affecting cell metabolism, bacterial adhesion, colonization, immunity, antibiotic resistance and serum resistance. The present study was an attempt to screen the marine environments like aquafarms and mangroves of coastal areas of Alappuzha and Cochin, Kerala for the presence of lysogenic V. cholerae, to study their pathogenicity and also gene transfer potential. Phenotypic and molecular methods were used for identification of isolates as V. cholerae. The thirty one isolates which were Gram negative, oxidase positive, fermentative, with or without gas production on MOF media and which showed yellow coloured colonies on TCBS (Thiosulfate Citrate Bile salt Sucrose) agar were segregated as vibrios. Twenty two environmental V. cholerae strains of both O1 and non- O1/non-O139 serogroups on induction with mitomycin C showed the presence of lysogenic phages. They produced characteristic turbid plaques in double agar overlay assay using the indicator strain V. cholerae El Tor MAK 757. PCR based molecular typing with primers targeting specific conserved sequences in the bacterial genome, demonstrated genetic diversity among these lysogen containing non-O1 V. cholerae . Polymerase chain reaction was also employed as a rapid screening method to verify the presence of 9 virulence genes namely, ctxA, ctxB, ace, hlyA, toxR, zot,tcpA, ninT and nanH, using gene specific primers. The presence of tcpA gene in ALPVC3 was alarming, as it indicates the possibility of an epidemic by accepting the cholera. Differential induction studies used ΦALPVC3, ΦALPVC11, ΦALPVC12 and ΦEKM14, underlining the possibility of prophage induction in natural ecosystems, due to abiotic factors like antibiotics, pollutants, temperature and UV. The efficiency of induction of prophages varied considerably in response to the different induction agents. The growth curve of lysogenic V. cholerae used in the study drastically varied in the presence of strong prophage inducers like antibiotics and UV. Bacterial cell lysis was directly proportional to increase in phage number due to induction. Morphological characterization of vibriophages by Transmission Electron Microscopy revealed hexagonal heads for all the four phages. Vibriophage ΦALPVC3 exhibited isometric and contractile tails characteristic of family Myoviridae, while phages ΦALPVC11 and ΦALPVC12 demonstrated the typical hexagonal head and non-contractile tail of family Siphoviridae. ΦEKM14, the podophage was distinguished by short non-contractile tail and icosahedral head. This work demonstrated that environmental parameters can influence the viability and cell adsorption rates of V. cholerae phages. Adsorption studies showed 100% adsorption of ΦALPVC3 ΦALPVC11, ΦALPVC12 and ΦEKM14 after 25, 30, 40 and 35 minutes respectively. Exposure to high temperatures ranging from 50ºC to 100ºC drastically reduced phage viability. The optimum concentration of NaCl required for survival of vibriophages except ΦEKM14 was 0.5 M and that for ΦEKM14 was 1M NaCl. Survival of phage particles was maximum at pH 7-8. V. cholerae is assumed to have existed long before their human host and so the pathogenic clones may have evolved from aquatic forms which later colonized the human intestine by progressive acquisition of genes. This is supported by the fact that the vast majority of V. cholerae strains are still part of the natural aquatic environment. CTXΦ has played a critical role in the evolution of the pathogenicity of V. cholerae as it can transmit the ctxAB gene. The unusual transformation of V. cholerae strains associated with epidemics and the emergence of V. cholera O139 demonstrates the evolutionary success of the organism in attaining greater fitness. Genetic changes in pathogenic V. cholerae constitute a natural process for developing immunity within an endemically infected population. The alternative hosts and lysogenic environmental V. cholerae strains may potentially act as cofactors in promoting cholera phage ‘‘blooms’’ within aquatic environments, thereby influencing transmission of phage sensitive, pathogenic V. cholerae strains by aquatic vehicles. Differential induction of the phages is a clear indication of the impact of environmental pollution and global changes on phage induction. The development of molecular biology techniques offered an accessible gateway for investigating the molecular events leading to genetic diversity in the marine environment. Using nucleic acids as targets, the methods of fingerprinting like ERIC PCR and BOX PCR, revealed that the marine environment harbours potentially pathogenic group of bacteria with genetic diversity. The distribution of virulence associated genes in the environmental isolates of V. cholerae provides tangible material for further investigation. Nucleotide and protein sequence analysis alongwith protein structure prediction aids in better understanding of the variation inalleles of same gene in different ecological niche and its impact on the protein structure for attaining greater fitness of pathogens. The evidences of the co-evolution of virulence genes in toxigenic V. cholerae O1 from different lineages of environmental non-O1 strains is alarming. Transduction studies would indicate that the phenomenon of acquisition of these virulence genes by lateral gene transfer, although rare, is not quite uncommon amongst non-O1/non-O139 V. cholerae and it has a key role in diversification. All these considerations justify the need for an integrated approach towards the development of an effective surveillance system to monitor evolution of V. cholerae strains with epidemic potential. Results presented in this study, if considered together with the mechanism proposed as above, would strongly suggest that the bacteriophage also intervenes as a variable in shaping the cholera bacterium, which cannot be ignored and hinting at imminent future epidemics.

Local food security and principle of organic farming (from farm to fork) in context of food culture in Indonesia: Minangkabau's case study

The basic idea behind improving local food security consists of two paths; first, accessibility (price, stock) and second, availability (quantity and biodiversity); both are perquisites to the provision of nutrients and a continuous food supply with locally available resources. The objectives of this thesis are to investigate if indigenous knowledge still plays an important role in traditional farming in the Minangkabau`s culture, thus supporting local food security. If the indigenous knowledge still plays a role in food culture in the Minangkabau`s culture which is linked to the matrilineal role and leads to a sound nutrition. Further, it should be tested if marantau influences traditional farming and food culture in Minangkabau`s, and if the local government plays a role in changing of traditional farming systems and food culture. Furthermore this thesis wants to prove if education and gender are playing a role in changing traditional farming system and food culture, and if the mass media affects traditional farming systems and food culture for the Minangkabau. The study was completed at four locations in West Sumatera; Nagari Ulakan (NU) (coastal area), Nagari Aia Batumbuak (NAB) (hilly area), Nagari Padang Laweh Malalo (NPLM) (lake area), Nagari Pandai Sikek (NPS) (hilly area). The rainfall ranged from 1400- 4800 mm annually with fertile soils. Data was collected by using PRA (Participatory Rural Appraisal) to investigate indigenous knowledge (IK) and its interactions, which is also combining with in depth-interview, life history, a survey using semi-structured-questionnaire, pictures, mapping, and expert interview. The data was collected from June - September 2009 and June 2010. The materials are; map of area, list of names, questionnaires, voices recorder, note book, and digital camera. The sampling method was snowball sampling which resulted in the qualitative and quantitative data taken. For qualitative data, ethnography and life history was used. For quantitative, a statistical survey with a semi-structured questionnaire was used. 50 respondents per each site participated voluntarily. Data was analyzed by performing MAXQDA 10, and F4 audio analysis software (created and developed by Philip-University Marburg). The data is clustered based on causality. The results show that; the role of IK on TFS (traditional farming system) shown on NPLM which has higher food crop biodiversity in comparison to the other three places even though it has relatively similar temperature and rainfall. This high food crop biodiversity is due to the awareness of local people who realized that they lived in unfavourable climate and topography; therefore they are more prepared for any changes that may occur. Carbohydrate intake is 100 % through rice even though they are growing different staple crops. Whereas most of the people said in the interviews that not eating rice is like not really eating for them. In addition to that, mothers still play an important role in kitchen activities. But when the agriculture income is low, mothers have to decide whether to change the meals or to feel insecure about their food supply. Marantau yields positive impact through the remittances it provides to invest on the farm. On the other hand, it results in fewer workers for agriculture, and therefore a negative impact on the transfer of IK. The investigation showed that the local government has a PTS (Padi Tanam Sabatang) programme which still does not guarantee that the farmers are getting sufficient revenue from their land. The low agricultural income leads to situation of potential food insecurity. It is evident that education is equal among men and women, but in some cases women tend to leave school earlier because of arranged marriages or the distances of school from their homes. Men predominantly work in agriculture and fishing, while women work in the kitchen. In NAB, even though women work on farmland they earn less then men. Weaving (NPS) and kitchen activity is recognized as women’s work, which also supports the household income. Mass media is not yielding any changes in TFS and food culture in these days. The traditional farming system has changed because of intensive agricultural extension which has introduced new methods of agriculture for the last three decades (since the 1980’s). There is no evidence that they want to change any of their food habits because of the mass media despite the lapau activity which allows them to get more food choices, instead preparing traditional meal at home. The recommendations of this thesis are: 1) The empowerment of farmers. It is regarding the self sufficient supply of manure, cooperative seed, and sustainable farm management. Farmers should know – where are they in their state of knowledge – so they can use their local wisdom and still collaborate with new sources of knowledge. Farmers should learn the prognosis of supply and demand next prior to harvest. There is a need for farm management guidelines; that can be adopted from both their local wisdom and modern knowledge. 2) Increase of non-agricultural income Increasing the non-agricultural income is strongly recommended. The remittances can be invested on non-agricultural jobs. 3) The empowerment of the mother. The mother plays an important role in farm to fork activities; the mother can be an initiator and promoter of cultivating spices in the backyard. Improvement of nutritional knowledge through information and informal public education can be done through arisan ibu-ibu and lapau activity. The challenges to apply these recommendations are: 1) The gap between institutions and organizations of local governments. There is more than one institution involved in food security policy. 2) Training and facilities for field extension agriculture (FEA) is needed because the rapid change of interaction between local government and farmer’s dependent on this agency.

Attitudes and behaviour towards food attributes and organic food – a triangular methodological investigation of occasional organic consumers

Organic food is increasingly available in the conventional food retail, where organic products are offered alongside with various other types of products and compete mainly with conventional and the so-called conventional-plus products. The latter are conventional products displaying particular quality attributes on the product packaging, such as ‘no artificial additives’, or ‘from animal welfare husbandry’. Often, these quality attributes also apply to organic products. Occasional organic consumers might prefer such conventional-plus alternatives that are perceived to be ‘between’ organic and conventional products. The overall objective of this PhD thesis was to provide information about the segment of occasional organic consumers. In particular, the thesis focussed on consumer perceptions and attitudes towards the quality of, and preferences for, organic, conventional and conventional-plus products in two countries: Germany and Switzerland. To achieve these objectives, qualitative and quantitative consumer research was combined in order to explore occasional organic consumers’ perceptions and attitudes as well as to observe their preferences and buying behaviour regarding different types of food products: organic, conventional and conventional-plus products. The qualitative research showed that, depending on single criteria, organic production was both positively as well as negatively assessed by consumers. Consumer perception of organic food was found to be highly selective and primarily focussed on the final stage of the particular production process. A major problem is that consumers are still mostly unfamiliar with factors associated with organic production, have a lack of confidence, and often confuse organic with conventional products. Besides this, consumer expectations of organic products are different from the expectations of conventional products. The quantitative research revealed that attitudes strongly determine consumers’ preferences for organic, conventional and conventional-plus products. Consumer attitudes tended to differ more between organic and conventional choices rather than conventional-plus and conventional choices. Furthermore, occasional organic consumers are heterogeneous in their preferences. They can be grouped into two segments: the consumers in one segment were less price sensitive and preferred organic products. The consumers in the other segment were more price sensitive and rather preferred conventional-plus or conventional products. To conclude, given the selective and subjective nature of consumer perception and the strong focus of consumer perception on the final stage of the food production process, specific additional values of organic farming should be communicated in clear and catchy messages. At the same time, these messages should be particularly focussed on the final stage of organic food production. The communication of specific added values in relation with organic products to improve the perceived price-performance-ratio is important since conventional-plus products represent an interesting alternative particularly for price sensitive occasional organic consumers. Besides this, it is important to strengthen affirmative consumer attitudes towards organic production. Therefore, policy support should emphasise on long-term communication campaigns and education programmes to increase the consumer awareness and knowledge of organic food and farming. Since consumers expect that organic food is regionally or at least domestically produced while they less accept organic imports, policy support of domestic and regional producers is a crucial measure to fill the current gap between the increasing consumer demand of organic food and the stagnation of the domestic and regional organic food supply.

Right to Food, Food Security and Food Aid Under International Law, or the Limits of a right-based approach

The right to food has become a pillar of international humanitarian and human rights law. The increasing number of food-related emergencies and the evolution of the international order brought the more precise notion of food security and made a potential right to receive food aid emerge. Despite this apparent centrality, recent statistics show that a life free from hunger is for many people all over the world still a utopian idea. The paper will explore nature and content of the right to food, food security and food aid under international law in order to understand the reasons behind the substantial failure of this right-centred approach, emphasising the lack of legal effects of many food-related provisions because of excessive moral connotations of the right to be free from hunger. Bearing in mind the three-dimensional nature of food security, the paper will also suggest that all attention has been focused on the availability of food, while real difficulties arise in terms of accessibility and adequacy. Emergency situations provide an excellent example of this unbalance, as the emerging right to receive food aid focus itself on the availability of food, without improving local production and adequacy. Looking at other evolving sectors of international law, such as the protection of the environment, and particularly the safeguard of biological diversity, alternative solutions will be envisaged in order to “feed” the right to food.

Connecting the Water and Carbon Cycles for the Generation of Food Security and Ecosystem Services

Water scarcity and food insecurity are pervasive issues in the developing world and are also intrinsically linked to one another. Through the connection of the water cycle and the carbon cycle this study illustrates that synergistic benefits can be realized by small scale farmers through the implementation of waste water irrigated agroforestry. The WaNuLCAS model is employed using La Huerta agroforestry site in Texcoco, South Central Mexico, as the basis for parameterization. The results of model simulations depicting scenarios of water scarcity and waste water irrigation clearly show that the addition of waste water greatly increases the agroforestry system’s generation of crop yields, above- and below-ground biomass, soil organic matter and carbon storage potential. This increase in carbon sequestration by the system translates into better local food security, diversified household income through payments for ecosystem services and contributes to the mitigation of global climate change.

From Market- to Development Orientation - The Trade Aspect of Food Security and Agriculture

Trade rules are suggested to be one of the reasons for the hunger in the world and environmental damage. As current trade rules encourage market orientation and therefore specialization and industrialization of agriculture, which has as side effects rural hunger and environmental damage, there is room for improvement in the international trade regime. One main finding of Nexus Foundations' work in Geneva is a possible new orientation for agricultural and food markets – an orientation on development, rather than purely on markets. This development orientation consists of several elements from development of soil fertility to local markets and consumer relatedness. Since the Bali Ministerial in 2013, the WTO has set up a four year work programme on the issue of food security related to food reserves. This opens the chance to discuss broader food security issues in the realm of trade negotiations.

Healthy food : Fruits and vegetables. 'Comida sana : frutas y verduras'.

Se trata de una unidad didáctica que pretende dar un conocimiento al alumno sobre un vocabulario técnico sobre frutas y verduras, con una puesta al día sobre los conocimientos previos del alumno se le darán unos conocimientos además de unas pautas alimentarias sobre alimentación saludable.

Edexcel 360 Science. Teacher pack : for GCSE biology, chemistry and physics.

Prepara a los estudiantes para la obtención del título General Certificate Secondary Education (GCSE) otorgado por el organismo Edexcel. Este recurso proporciona al profesor las respuestas a las preguntas del libro del alumno, apoyo adicional con cuadernos de trabajo y actividades prácticas para las clases. Al texto impreso le acompaña un CD-ROM con material didáctico en formato Word y PDF. Tiene como material de apoyo el libro del alumno.

Edexcel 360science. Students' book extension units for GCSE Biology, GCSE Chemistry and GCSE Physics : Edexcel's own course for the new specification.

El objetivo de este recurso es involucrar y estimular a los alumnos a conocer cómo funciona la ciencia y les ayuda a adquirir habilidades prácticas y capacidades analíticas básicas para desarrollar las preguntas de evaluación. La publicación está dividida en seis bloques, cada uno cuenta con dieciocho secciones de dos páginas. Al final de cada bloque hay un conjunto de preguntas para practicar los exámenes y glosario de palabras clave. Incluye CD-ROM.

OCR Home Economics for A2: Food, nutrition and health today.

Este libro está diseñado para apoyar a los estudiantes en el estudio y preparación de la asignatura economía doméstica para OCR nivel A2 de enseñanza secundaria. Los temas del libro son: introducción a la nutrición (concepto de nutrición, alimentación equilibrada, malnutrición), los nutrientes y la energía (las proteínas, las grasas, los hidratos de carbono, los minerales: calcio, potasio, sodio, las fuentes de energía), necesidades nutricionales y dietéticas de los diferentes grupos (niños, adolescentes, adultos, vegetarianos, mujeres embarazadas), propiedades de la comida, diseño, desarrollo y producción de nuevos productos.