Data analysis tools and methods for DNA microarray and high-throughput sequencing data

The recent rapid development of biotechnological approaches has enabled the production of large whole genome level biological data sets. In order to handle thesedata sets, reliable and efficient automated tools and methods for data processingand result interpretation are required. Bioinformatics, as the field of studying andprocessing biological data, tries to answer this need by combining methods and approaches across computer science, statistics, mathematics and engineering to studyand process biological data. The need is also increasing for tools that can be used by the biological researchers themselves who may not have a strong statistical or computational background, which requires creating tools and pipelines with intuitive user interfaces, robust analysis workflows and strong emphasis on result reportingand visualization. Within this thesis, several data analysis tools and methods have been developed for analyzing high-throughput biological data sets. These approaches, coveringseveral aspects of high-throughput data analysis, are specifically aimed for gene expression and genotyping data although in principle they are suitable for analyzing other data types as well. Coherent handling of the data across the various data analysis steps is highly important in order to ensure robust and reliable results. Thus,robust data analysis workflows are also described, putting the developed tools andmethods into a wider context. The choice of the correct analysis method may also depend on the properties of the specific data setandthereforeguidelinesforchoosing an optimal method are given. The data analysis tools, methods and workflows developed within this thesis have been applied to several research studies, of which two representative examplesare included in the thesis. The first study focuses on spermatogenesis in murinetestis and the second one examines cell lineage specification in mouse embryonicstem cells.

Catching up with competitiveness in emerging markets – An analysis of the role of the firm’s technology management strategies

Emerging markets have experienced rapid economic growth, and manufacturing firms have had to face the effects of globalisation. Some of the major emerging economies have been able to create a supportive business environment that fosters innovation, and China is a good example of a country that has been able to increase value-added investments. Conversely, when we look at Russia, another big emerging market, we witness a situation in which domestic firms struggle more with global competitiveness. Innovation has proven to be one of the most essential ingredients for firms aiming to grow and become more competitive. In emerging markets, the business environment sets many constraints for innovation. However, open strategic choices in new product development enable companies in emerging markets to expand their resource base and capability building. Networking and close inter-firm cooperation are essential in this regard. In this dissertation, I argue that technology transfer is one of the key tools for these companies to become internationally networked and to improve their competitiveness. It forces companies to reach outside the company and national borders, which in many cases, is a major challenge for firms in emerging markets. This dissertation focuses on how companies can catch up with competitiveness in emerging markets. The empirical studies included in the dissertation are based on analyses of survey data mainly of firms and their strategies in the Russian manufacturing industry. The dissertation contributes to the current strategic management literature by further investigating technology management strategies in manufacturing firms in emerging markets and the benefits of more open approaches to new product development and innovation.

Understanding the bioactivity of plant tannins: developments in analysis methods and structure-activity studies

Tannins, typically segregated into two major groups, the hydrolyzable tannins (HTs) and the proanthocyanidins (PAs), are plant polyphenolic secondary metabolites found throughout the plant kingdom. On one hand, tannins may cause harmful nutritional effects on herbivores, for example insects, and hence they work as plants’ defense against plant-eating animals. On the other hand, they may affect positively some herbivores, such as mammals, for example by their antioxidant, antimicrobial, anti-inflammatory or anticarcinogenic activities. This thesis focuses on understanding the bioactivity of plant tannins, their anthelmintic properties and the tools used for the qualitative and quantitative analysis of this endless source of structural diversity. The first part of the experimental work focused on the development of ultra-high performance liquid chromatography−tandem mass spectrometry (UHPLC-MS/MS) based methods for the rapid fingerprint analysis of bioactive polyphenols, especially tannins. In the second part of the experimental work the in vitro activity of isolated and purified HTs and their hydrolysis product, gallic acid, was tested against egg hatching and larval motility of two larval developmental stages, L1 and L2, of a common ruminant gastrointestinal parasite, Haemonchus contortus. The results indicated clear relationships between the HT structure and the anthelmintic activity. The activity of the studied compounds depended on many structural features, including size, functional groups present in the structure, and the structural rigidness. To further understand tannin bioactivity on a molecular level, the interaction between bovine serum albumin (BSA), and seven HTs and epigallocatechin gallate was examined. The objective was to define the effect of pH on the formation on tannin–protein complexes and to evaluate the stability of the formed complexes by gel electrophoresis and MALDI-TOF-MS. The results indicated that more basic pH values had a stabilizing effect on the tannin–protein complexes and that the tannin oxidative activity was directly linked with their tendency to form covalently stabilized complexes with BSA at increased pH.

An analysis of students' travel motivations and images of China as a tourist destination

Despite China's rapid growth in inbound tourism, the nature of its Canadian tourist market has been insufficiently studied. In response to this need, the objectives of this study are to identify China's destination image in Canadian students' minds, their possible internal motivations for visiting China as well as examining demographic influences on people's destination image formation. The study reviews image formation process and travel motivation categorisation, discusses their relationship, and implements Baloglu and McCleary's (1999) perceptual and affective image formation model and "push and pull factors" theory as its framework. A self-administered survey was applied to 424 undergraduate students in a Canadian university in early 2004. Exploratory factor analyses were conducted to identify perceived images and travel motivation. Summated means were calculated to illustrate the affective attitudes. A series of f-test and ANOVA tests were employed to examine the influence of demographics. An open-ended question format was adopted to analyse other images, motivations and visitation barriers that students may have. Findings demonstrate that cultural and natural attractions are the predominant image which the Canadian students have of China'; some stereotypes and negative images still influence the students' perception; travel service quality is largely unknown; increasing knowledge and seeking excitement and fun are the significant motivators in the likelihood of the Canadian students choosing to visit China; and personal interests may be a factor that significantly influences an individual's destination image and travel motivation. Raising awareness and increasing familiarity through promotion are suggested as methods to create a positive destination image of China.

Isolation and analysis of a corprinus cinereus DNA fragment showing homology to a fimbrial cDNA from ustilago violacea

Surface proteinaceous fibrils, termed fimbriae, were first identified on gram negative bacteria in the 1940s. Fungal fimbriae, discovered some 25 years later, are found on members of all fungal classes. In the present study, polyclonal antiserum raised against the fimbrial proteins of U. vio/acea were used in order to identify antigenically related proteins from Coprinus cinereus and Schizophy//um commune. Two polypeptides with molecular masses of 37 and 39 kDa from C. cinereus were observed and confirm earlier results. A single previously unidentified 50 kDa polypeptide in S. commune crossreacted with the antiserum. The 50 kDa protein was found to consist of 3 isoforms with isoelectric points ranging from 5.6 to 5.8. A fimbrial cDNA derived from U. vio/acea was used to identify DNA restriction fragments from C. cinereus and S. commune showing homology to the fimbrial transcript of U. vio/acea. Heterologous hybridization with this cDNA was used in order to screen a C. cinereus genomic DNA library. A single clone, A2-3A, with a 14 kbp insert showed strong homology to the pfim3-1 cDNA. The region of homology, a 700 bp Xba I fragment, was subcloned into pUG19. This plasmid was refered to as pXX8. DNA sequence determinations of pXX8 and adjacent fragments from A2-3A suggested that the cloned DNA was a portion of the rONA repeat encoding the small subunit rRNA. DNA sequence analysis of pfim3-1 yielded an incomplete open reading frame. The predicted amino acid sequence codes for a 206 amino acid, 22 kDa polypeptide which contains a domain similar to a transmembrane domain from rat leukocyte antigen, GDS3. As well, an untranslated 576 nucleotide domain showed 81 % homology to pXX8 and 830/0 homology to the 188 rRNA sequence of Ustilago maydis. This sequence was found adjacent to a region of adenine-thymine base pairs presumed to represent the polyadenylation sequence of the fimbrial transcript. The size and extent of homology is sufficient to account for the hybridization of pfim3-1 to rDNA. It is suggested that this domain represents a completely novel regulatory domain within eukaryotes that may enable the observed rapid regeneration of fimbriae in U. violacea.

Sustainable Solid Waste Management Through Ecological Footprint Analysis – A Solution To Kochi City, India

Kochi, the commercial capital of Kerala, South India and second most important city next to Mumbai on the Western coast is a land having a wide variety of residential environments. Due to rapid population growth, changing lifestyles, food habits and living standards, institutional weaknesses, improper choice of technology and public apathy, the present pattern of the city can be classified as that of haphazard growth with typical problems characteristics of unplanned urban development especially in the case of solid waste management. To have a better living condition for us and our future generations, we must know where we are now and how far we need to go. We, each individual must calculate how much nature we use and compare it to how much nature we have available. This can be achieved by applying the concept of ecological footprint. Ecological footprint analysis (EFA) is a quantitative tool that represents the ecological load imposed on earth by humans in spatial terms. The aim of applying EFA to Kochi city is to quantify the consumption and waste generation of a population and to compare it with the existing biocapacity. By quantifying the ecological footprint we can formulate strategies to reduce the footprint and there by having a sustainable living. The paper discusses the various footprint components of Kochi city and in detail analyses the waste footprint of the residential areas using waste footprint analyzer. An attempt is also made to suggest some waste foot print reduction strategies thereby making the city sustainable as far as solid waste management is concerned.

Sustainable Solid Waste Management Solutions To Kochi City, India, Through The Environmental Management Tool Ecological Footprint Analysis

Solid waste management nowadays is an important environmental issue in country like India. Statistics show that there has been substantial increase in the solid waste generation especially in the urban areas. This trend can be ascribed to rapid population growth, changing lifestyles, food habits, and change in living standards, lack of financial resources, institutional weaknesses, improper choice of technology and public apathy towards municipal solid waste. Waste is directly related to the consumption of resources and dumping to the land. Ecological footprint analysis – an impact assessment environment management tool makes a relationship between two factors- the amount of land required to dispose per capita generated waste. Ecological footprint analysis is a quantitative tool that represents the ecological load imposed on the earth by humans in spatial terms. By quantifying the ecological footprint we can formulate strategies to reduce the footprint and there by having a sustainable living. In this paper, an attempt is made to explore the tool Ecological Footprint Analysis with special emphasis to waste generation. The paper also discusses and analyses the waste footprint of Kochi city,India. An attempt is also made to suggest strategies to reduce the waste footprint thereby making the city sustainable, greener and cleaner

Ecological Foot Print Analysis-A Sustainable Environmental Management Tool for Kochi City

Kochi, the commercial capital of Kerala and the second most important city next to Mumbai on the Western coast of India, is a land having a wide variety of residential environments. The present pattern of the city can be classified as that of haphazard growth with typical problems characteristics of unplanned urban development. This trend can be ascribed to rapid population growth, our changing lifestyles, food habits, and change in living standards, institutional weaknesses, improper choice of technology and public apathy. Ecological footprint analysis (EFA) is a quantitative tool that represents the ecological load imposed on the earth by humans in spatial terms. This paper analyses the scope of EFA as a sustainable environmental management tool for Kochi City

Sustainable Development through the Environment Management Tool Ecological Footprint Analysis – A Study in Kochi City, India

In the past, natural resources were plentiful and people were scarce. But the situation is rapidly reversing. Our challenge is to find a way to balance human consumption and nature’s limited productivity in order to ensure that our communities are sustainable locally, regionally and globally. Kochi, the commercial capital of Kerala, South India and the second most important city next to Mumbai on the Western coast is a land having a wide variety of residential environments. Due to rapid population growth, changing lifestyles, food habits and living standards, institutional weaknesses, improper choice of technology and public apathy, the present pattern of the city can be classified as that of haphazard growth with typical problems characteristics of unplanned urban development. Ecological Footprint Analysis (EFA) is physical accounting method, developed by William Rees and M. Wackernagel, focusing on land appropriation using land as its “currency”. It provides a means for measuring and communicating human induced environmental impacts upon the planet. The aim of applying EFA to Kochi city is to quantify the consumption and waste generation of a population and to compare it with the existing biocapacity. By quantifying the ecological footprint we can formulate strategies to reduce the footprint and there by having a sustainable living. In this paper, an attempt is made to explore the tool Ecological Footprint Analysis and calculate and analyse the ecological footprint of the residential areas of Kochi city. The paper also discusses and analyses the waste footprint of the city. An attempt is also made to suggest strategies to reduce the footprint thereby making the city sustainable

Export performancecof Indian spices in the WTO regime: a disaggregated analysis

Export has assumed an important place in the development of any country and considered as the engine of economic growth. India requires huge amount of foreign exchange for its essential import and for achieving rapid growth. Millions of job opportunities have to be created to utilise the youth for nation building. Even though the country has different sources of foreign exchange, export earning is the safe way of obtaining it in the long run. Export of high valued traditional products not only gives foreign exchange, but also employment to large number of people. Spices are the traditional products of India whose production process is highly intensive in semi and unskilled labour, and high domestic and foreign market prices compared to other traditional products. The new world trade scenario with the establishment of WTO has affected India’s spices export considerably. The study examines the export performance of Indian spices in the WTO regime taking the export of major spices from 1985 to 2013 using the growth of export, trend and instability in growth rate, changes in the composition and direction of spices, export performance ratio and the prospects of spices in earning foreign exchange during the WTO period and Pre-WTO period. The analysis reveals that the overall performance of Indian spices exports during the WTO regime are satisfactory. Export volume and value increased much during this period. But the decrease in market share of spices export during the WTO period reflects that, the favourable conditions in the international market are not exploited by India. High Revealed Comparative Advantage (RCA) and Value Elasticity (EV) of major spices amidst the low export shares shows that export performance of Indian spices during the WTO regime was not mainly affected by external demand factors as suggested by Ragnar Nurkse in his Demand Deficiency Thesis, but because of internal supply factors as suggested in Supply Deficiency Thesis, (supported by K.S Dhinsha, Dacosta, Goddamwar,etc.). But the fluctuations of export during the recession and prosperity periods show that external demand is also a determinant of Indian spices export. From this one can conclude that both the domestic supply factors and foreign demand factors influence the export performance of Indian spices. The long term performance of Indian spices exports are mainly influenced by domestic supply factors as suggested by Supply Deficiency Thesis and short term performance is mostly influenced by external demand factors as suggested by Demand Deficiency Thesis.

A Modeling Analysis of Changing Global Land Use as affected by Changing Demands for Wood Products

The 21st century has brought new challenges for forest management at a time when globalization in world trade is increasing and global climate change is becoming increasingly apparent. In addition to various goods and services like food, feed, timber or biofuels being provided to humans, forest ecosystems are a large store of terrestrial carbon and account for a major part of the carbon exchange between the atmosphere and the land surface. Depending on the stage of the ecosystems and/or management regimes, forests can be either sinks, or sources of carbon. At the global scale, rapid economic development and a growing world population have raised much concern over the use of natural resources, especially forest resources. The challenging question is how can the global demands for forest commodities be satisfied in an increasingly globalised economy, and where could they potentially be produced? For this purpose, wood demand estimates need to be integrated in a framework, which is able to adequately handle the competition for land between major land-use options such as residential land or agricultural land. This thesis is organised in accordance with the requirements to integrate the simulation of forest changes based on wood extraction in an existing framework for global land-use modelling called LandSHIFT. Accordingly, the following neuralgic points for research have been identified: (1) a review of existing global-scale economic forest sector models (2) simulation of global wood production under selected scenarios (3) simulation of global vegetation carbon yields and (4) the implementation of a land-use allocation procedure to simulate the impact of wood extraction on forest land-cover. Modelling the spatial dynamics of forests on the global scale requires two important inputs: (1) simulated long-term wood demand data to determine future roundwood harvests in each country and (2) the changes in the spatial distribution of woody biomass stocks to determine how much of the resource is available to satisfy the simulated wood demands. First, three global timber market models are reviewed and compared in order to select a suitable economic model to generate wood demand scenario data for the forest sector in LandSHIFT. The comparison indicates that the ‘Global Forest Products Model’ (GFPM) is most suitable for obtaining projections on future roundwood harvests for further study with the LandSHIFT forest sector. Accordingly, the GFPM is adapted and applied to simulate wood demands for the global forestry sector conditional on selected scenarios from the Millennium Ecosystem Assessment and the Global Environmental Outlook until 2050. Secondly, the Lund-Potsdam-Jena (LPJ) dynamic global vegetation model is utilized to simulate the change in potential vegetation carbon stocks for the forested locations in LandSHIFT. The LPJ data is used in collaboration with spatially explicit forest inventory data on aboveground biomass to allocate the demands for raw forest products and identify locations of deforestation. Using the previous results as an input, a methodology to simulate the spatial dynamics of forests based on wood extraction is developed within the LandSHIFT framework. The land-use allocation procedure specified in the module translates the country level demands for forest products into woody biomass requirements for forest areas, and allocates these on a five arc minute grid. In a first version, the model assumes only actual conditions through the entire study period and does not explicitly address forest age structure. Although the module is in a very preliminary stage of development, it already captures the effects of important drivers of land-use change like cropland and urban expansion. As a first plausibility test, the module performance is tested under three forest management scenarios. The module succeeds in responding to changing inputs in an expected and consistent manner. The entire methodology is applied in an exemplary scenario analysis for India. A couple of future research priorities need to be addressed, particularly the incorporation of plantation establishments; issue of age structure dynamics; as well as the implementation of a new technology change factor in the GFPM which can allow the specification of substituting raw wood products (especially fuelwood) by other non-wood products.

Digital image analysis as a tool to estimate legume contributions in legume-grass swards

Summary: Productivity and forage quality of legume-grass swards are important factors for successful arable farming in both organic and conventional farming systems. For these objectives the botanical composition of the swards is of particular importance, especially, the content of legumes due to their ability to fix airborne nitrogen. As it can vary considerably within a field, a non-destructive detection method while doing other tasks would facilitate a more targeted sward management and could predict the nitrogen supply of the soil for the subsequent crop. This study was undertaken to explore the potential of digital image analysis (DIA) for a non destructive prediction of legume dry matter (DM) contribution of legume-grass mixtures. For this purpose an experiment was conducted in a greenhouse, comprising a sample size of 64 experimental swards such as pure swards of red clover (Trifolium pratense L.), white clover (Trifolium repens L.) and lucerne (Medicago sativa L.) as well as binary mixtures of each legume with perennial ryegrass (Lolium perenne L.). Growth stages ranged from tillering to heading and the proportion of legumes from 0 to 80 %. Based on digital sward images three steps were considered in order to estimate the legume contribution (% of DM): i) The development of a digital image analysis (DIA) procedure in order to estimate legume coverage (% of area). ii) The description of the relationship between legume coverage (% area) and legume contribution (% of DM) derived from digital analysis of legume coverage related to the green area in a digital image. iii) The estimation of the legume DM contribution with the findings of i) and ii). i) In order to evaluate the most suitable approach for the estimation of legume coverage by means of DIA different tools were tested. Morphological operators such as erode and dilate support the differentiation of objects of different shape by shrinking and dilating objects (Soille, 1999). When applied to digital images of legume-grass mixtures thin grass leaves were removed whereas rounder clover leaves were left. After this process legume leaves were identified by threshold segmentation. The segmentation of greyscale images turned out to be not applicable since the segmentation between legumes and bare soil failed. The advanced procedure comprising morphological operators and HSL colour information could determine bare soil areas in young and open swards very accurately. Also legume specific HSL thresholds allowed for precise estimations of legume coverage across a wide range from 11.8 - 72.4 %. Based on this legume specific DIA procedure estimated legume coverage showed good correlations with the measured values across the whole range of sward ages (R2 0.96, SE 4.7 %). A wide range of form parameters (i.e. size, breadth, rectangularity, and circularity of areas) was tested across all sward types, but none did improve prediction accuracy of legume coverage significantly. ii) Using measured reference data of legume coverage and contribution, in a first approach a common relationship based on all three legumes and sward ages of 35, 49 and 63 days was found with R2 0.90. This relationship was improved by a legume-specific approach of only 49- and 63-d old swards (R2 0.94, 0.96 and 0.97 for red clover, white clover, and lucerne, respectively) since differing structural attributes of the legume species influence the relationship between these two parameters. In a second approach biomass was included in the model in order to allow for different structures of swards of different ages. Hence, a model was developed, providing a close look on the relationship between legume coverage in binary legume-ryegrass communities and the legume contribution: At the same level of legume coverage, legume contribution decreased with increased total biomass. This phenomenon may be caused by more non-leguminous biomass covered by legume leaves at high levels of total biomass. Additionally, values of legume contribution and coverage were transformed to the logit-scale in order to avoid problems with heteroscedasticity and negative predictions. The resulting relationships between the measured legume contribution and the calculated legume contribution indicated a high model accuracy for all legume species (R2 0.93, 0.97, 0.98 with SE 4.81, 3.22, 3.07 % of DM for red clover, white clover, and lucerne swards, respectively). The validation of the model by using digital images collected over field grown swards with biomass ranges considering the scope of the model shows, that the model is able to predict legume contribution for most common legume-grass swards (Frame, 1992; Ledgard and Steele, 1992; Loges, 1998). iii) An advanced procedure for the determination of legume DM contribution by DIA is suggested, which comprises the inclusion of morphological operators and HSL colour information in the analysis of images and which applies an advanced function to predict legume DM contribution from legume coverage by considering total sward biomass. Low residuals between measured and calculated values of legume dry matter contribution were found for the separate legume species (R2 0.90, 0.94, 0.93 with SE 5.89, 4.31, 5.52 % of DM for red clover, white clover, and lucerne swards, respectively). The introduced DIA procedure provides a rapid and precise estimation of legume DM contribution for different legume species across a wide range of sward ages. Further research is needed in order to adapt the procedure to field scale, dealing with differing light effects and potentially higher swards. The integration of total biomass into the model for determining legume contribution does not necessarily reduce its applicability in practice as a combined estimation of total biomass and legume coverage by field spectroscopy (Biewer et al. 2009) and DIA, respectively, may allow for an accurate prediction of the legume contribution in legume-grass mixtures.

Value Stream Analysis and Mapping for Product Development

This paper explores the concept of Value Stream Analysis and Mapping (VSA/M) as applied to Product Development (PD) efforts. Value Stream Analysis and Mapping is a method of business process improvement. The application of VSA/M began in the manufacturing community. PD efforts provide a different setting for the use of VSA/M. Site visits were made to nine major U.S. aerospace organizations. Interviews, discussions, and participatory events were used to gather data on (1) the sophistication of the tools used in PD process improvement efforts, (2) the lean context of the use of the tools, and (3) success of the efforts. It was found that all three factors were strongly correlated, suggesting success depends on both good tools and lean context. Finally, a general VSA/M method for PD activities is proposed. The method uses modified process mapping tools to analyze and improve process.

Analysis of Pleistocene paleodrainage evolution in the Po Basin (Italy) by multivariate statistical techniques

In order to obtain a high-resolution Pleistocene stratigraphy, eleven continuously cored boreholes, 100 to 220m deep were drilled in the northern part of the Po Plain by Regione Lombardia in the last five years. Quantitative provenance analysis (QPA, Weltje and von Eynatten, 2004) of Pleistocene sands was carried out by using multivariate statistical analysis (principal component analysis, PCA, and similarity analysis) on an integrated data set, including high-resolution bulk petrography and heavy-mineral analyses on Pleistocene sands and of 250 major and minor modern rivers draining the southern flank of the Alps from West to East (Garzanti et al, 2004; 2006). Prior to the onset of major Alpine glaciations, metamorphic and quartzofeldspathic detritus from the Western and Central Alps was carried from the axial belt to the Po basin longitudinally parallel to the SouthAlpine belt by a trunk river (Vezzoli and Garzanti, 2008). This scenario rapidly changed during the marine isotope stage 22 (0.87 Ma), with the onset of the first major Pleistocene glaciation in the Alps (Muttoni et al, 2003). PCA and similarity analysis from core samples show that the longitudinal trunk river at this time was shifted southward by the rapid southward and westward progradation of transverse alluvial river systems fed from the Central and Southern Alps. Sediments were transported southward by braided river systems as well as glacial sediments transported by Alpine valley glaciers invaded the alluvial plain. Kew words: Detrital modes; Modern sands; Provenance; Principal Components Analysis; Similarity, Canberra Distance; palaeodrainage

Analysis of water-level response to rainfall and implications for recharge pathways in the Chalk aquifer, SE England

Water table response to rainfall was investigated at six sites in the Upper, Middle and Lower Chalk of southern England. Daily time series of rainfall and borehole water level were cross-corretated to investigate seasonal variations in groundwater-level response times, based on periods of 3-month duration. The time tags (in days) yielding significant correlations were compared with the average unsaturated zone thickness during each 3-month period. In general, for cases when the unsaturated zone was greater than 18 m thick, the time tag for a significant water-level response increased rapidly once the depth to the water table exceeded a critical value, which varied from site to site. For shallower water tables, a linear relationship between the depth to the water table and the water-level response time was evident. The observed variations in response time can only be partially accounted for using a diffusive model for propagation through the unsaturated matrix, suggesting that some fissure flow was occurring. The majority of rapid responses were observed during the winter/spring recharge period, when the unsaturated zone is thinnest and the unsaturated zone moisture content is highest, and were more likely to occur when the rainfall intensity exceeded 5 mm/day. At some sites, a very rapid response within 24 h of rainfall was observed in addition to the longer term responses even when the unsaturated zone was up to 64 m thick. This response was generally associated with the autumn period. The results of the cross-correlation analysis provide statistical support for the presence of fissure flow and for the contribution of multiple pathways through the unsaturated zone to groundwater recharge. (c) 2006 Elsevier B.V. All rights reserved.