Food webs in the era of molecular revolution – Like resolving the Gordian knot with a tricorder

Living nature consists of countless organisms, which are classified into millions of species. These species interact in many ways; for example predators when foraging on their prey, insect larvae consuming plants, and pathogenic bacteria drifting into humans. In addition, abiotic nature has a great initiative impact on life through many factors (including sunlight, ambient temperature, and water. In my thesis, I have studied interactions among different life forms in multifaceted ways. The webs of these interactions are commonly referred to as food webs, describing feeding relationships between species or energy transfer from one trophic level to another. These ecological interactions – whether they occur between species, between individuals, or between microorganisms within an individual – are among the greatest forces affecting natural communities. Relationships are tightly related to biological diversity, that is, species richness and abundances. A species is called a node in food web vocabulary, and its interactions to other species are called links. Generally, Artic food webs are considered to be loosely linked, simple structures. This conception roots into early modern food webs, where insects and other arthropods, for example, were clumped under one node. However, it has been shown that arthropods form the greatest part of diversity and biomass both in the tropics and in Arctic areas. Earlier challenges of revealing the role of insects and microorganisms in interactions webs have become possible with the help of recent advances in molecular techniques. In the first chapter, I studied the prey diversity of a common bat, Myotis daubentonii, in southwestern Finland. My results proved M. daubentonii being a versatile predator whose diet mainly consists of aquatic insects, such as chironomid midges. In the second chapter, I expanded the view to changes in seasonal and individual-based variation in the diet of M. daubentonii including the relationship between available and observed prey. I found out that chironomids remain the major prey group even though their abundance decreases in proportion to other insect groups. Diet varied a lot between individuals, although the differences were not statistically significant. The third chapter took the study to a large network in Greenland. I showed that Artic food webs are very complex when arthropods are taken into account. In the fourth chapter, I examined the bacterial flora of M. daubentonii and surveyed the zoonotic potential of these bacteria. I found Bartonella bacteria, of which one was described as a new species named after the locality of discovery. I have shown in my thesis that Myotis daubentonii as a predator links many insect species as well as terrestrial and aquatic environments. Moreover, I have exposed that Arctic food webs are complex structures comprising of many densely linked species. Finally, I demonstrated that the bacterial flora of bats includes several previously unknown species, some of which could possibly turn in to zoonosis. To summarize, molecular methods have untied several knots in biological research. I hope that this kind of increasing knowledge of the surrounding nature makes us further value all the life forms on earth.

CCK, dietary variety, and food intake in rats /

Among the environmental factors that can affect food intake is the extent of dietary variety available in the environment. Numerous studies have demonstrated that variety in a meal can increase the amount of food consumed in humans, rats, and other species. A physiological mechanism that has been demonstrated to affect food intake is the gut peptide cholecystokinin (CCK) which is released from the upper small intestine during the ingestion of food. Peripherally administered CCK has a robust inhibitory effect on the intake of a single-food meal. Thus, dietary variety and CCK both affect meal size, with dietary variety increasing intake and CCK decreasing intake. This raises the question ofhow dietary variety and CCK might interact to affect meal size. Previous studies of CCK's effects have focused on situations in which only one food was available for consumption. However, in an animal's natural environment it would frequently occur that the animal would come across a number of foods either simultaneously or in quick succession, thus providing the animal access to a variety of foods during a meal. Accordingly, the effect ofCCK on food intake in single-food and multiple-food meals was examined. It was found that food intake was greater in multiple-food than in single-food meals provided that foods in the multiplefood meal were presented either simultaneously or in increasing order of preference. When foods in the multiple-food meal were presented in decreasing order of preference, intake was similar to that observed in single-food meals. In addition, it was found that CCK inhibited food intake in a dose-dependent manner, and that its effects on food intake were similar regardless of meal type. Therefore, the inhibitory effects ofCCK were not diminished when a variety of foods were available for consumption. Furthermore, the finding that CCK did not differentially affect the intake of the two types of meals does not provide support for the recent-foods hypothesis which postulates that CCK decreases food intake by reducing the palatability of only recently consumed foods. However, it is consistent with the all-foods hypothesis, which predicts that CCK reduces food intake by decreasing the palatability of all foods. The 600 ng/kg dose of the CCK^-antagonist lorglumide significantly antagonized the inhibitory effect of exogenous CCK on food intake, and the magnitude of this effect was similar for both types of meal. These results suggest that exogenous CCK inhibits food intake through the activation ofCCK^ receptors. However, when administered by itself, the 600^ig/kg dose of lorglumide did not increase food intake in either single-food or multiple-food meals, suggesting that peripheral endogenous CCK may not play a major role in the control of food intake.

Rheological characteristics of suwari and kamaboko gels made of surimi from Indian major carps

The gel strength, compressibility and folding characteristic of suwari (set) and kamaboko (set and cooked) gels prepared from rohu (Labeo rohita), catla (Catla catla) and mrigal (Cirrhinus mrigala) surimi were examined to understand the occurrence of suwari and modori phenomena in surimi from major freshwater carps. Suwari setting of gels did not take place at lower temperatures. Suwari gels showed good gel strength at 50 °C for rohu and at 60 °C for catla and mrigal after 30 min setting time. Incubation for 60 min decreased the gel strength at 60 °C for rohu and catla. Setting at 25 °C followed by cooking at 90 °C increased the gel strength. Increased setting temperature, however, decreased the gel strength of cooked gels. Gel strength and compressibility data were supported by folding characteristics.

Targeted Public Distribution System in Food Grains:Extent of Utlization by the Tribal Population in Kerala

In India, Food Security meant supply of food grains and the medium was Public Distribution System. Public Distribution System (PDS) is a rationing mechanism that entitles households to specified quantities of selected commodities at subsidized prices. The Objectives of PDS are maintaining Price Stability, rationing during times of scarcity, welfare of the poor, and keeping a check on private trade. Kerala has registered remarkable improvement in poverty reduction in general over the years among all social sections, including scheduled caste and scheduled tribe population. As part of the structural adjustment intended to reduce public expenditure, PDS has been modified as Revamped PDS (RPDS) during 1992 and later on as Targeted PDS (TPDS) in 1997, intended to target households on the basis of income criterion, classifying people as Below Poverty Line (BPL) and Above Poverty Line (APL). TPDS provides 25Kg. of food gra.ins through the Fair Price Shops per month @ Rs.3/- per Kg. of rice/ wheat to the BPL category and @Rs.8.90 and Rs.6.7O for rice and wheat respectively to the APL category of people. Since TPDS is intended to target the poor people, the subsidy spent by the government for the scheme should be beneficial to the poor people and naturally they should utilize the benefits by purchasing the food grains allotted under the scheme. Several studies have shown that there is underutilization of the allotments under TPDS. Therefore, the extent of utilization of TPDS in food grains, how and why remains as a major hurdle, in improving the structure and system of PDS. Livelihood of the tribal population being under threat due to increasing degradation of the resources, the targeting system ought to be effective among the tribal population. Therefore, performance of the TPDS in food grains, in terms of the utilization by the tribal population in Kerala, impact thereof and the factors, if any, affecting proper utilization were considered as the research problem in this study. The study concentrated on the pattern of consumption of food grains by the tribal people, whether their hunger needs are met by distribution of food grains through the TPDS, extent to which TPDS in food grains reduce their share of expenditure on food in the total household expenditure, and the factors affecting the utilization of the TPDS in food grains by the tribal population. Going through the literature, it has been noted that only few studies concentrated on the utilization of TPDS in food grains among the tribal population in Kerala.The Research Design used in this study is descriptive in nature, but exploratory in some aspects. Idukki, Palakkad and Wayanad have more than 60% of the population of the tribals in the state. Within the three districts mentioned above, 14 villages with scheduled tribe concentration were selected for the study. 95 tribal colonies were selected from among the various tribal settlements. Collection of primary data was made from 1231 households with in the above tribal colonies. Analysis of data on the socio-economic factors of the tribal people, pattern of food consumption, extent of reduction in the share of expenditure on food among the household expenditure of the tribal people and the impact of TPDS on the tribal families etc. and testing of hypotheses to find out the relation/association of each of the six variables, using the data on BPL and APL categories of households separately have resulted in findings such as six percent of the tribal families do not have Ration Cards, average per capita consumption of food grains by the tribal people utilizing TPDS meets 62% of their minimum requirement, whereas the per capita consumption of food grains by the tribal people is higher than the national average per capita consumption, 63% deficiency in food grains may be felt by tribal people in general, if TPDS is withdrawn, and the deficit for BPL tribal people may be 82%, TPDS facilitates a reduction of 9.71% in the food expenditure among the total household expenditure of the tribal people in general, share of food to non-food among BPL category of tribals is 55:45 and 40:60 among the APL, Variables, viz. household income, number of members in the family and distance of FPS from tribal settlements etc. have influence on the quantity of rice being purchased by the tribal people from the Fair Price Shops, and there is influence of household income and distance of FPS from tribal settlements on the quantity of rice being purchased by the tribal people from the open market. Rationing with differential pricing on phased allotments, rectification of errors in targeting, anomalies in norms and procedures for classifying tribal people as BPL/APL, exclusive Income Generation for tribal population, paddy cultivation in the landholdings possessed by the tribal people, special drive for allotment of Ration Cards to the tribal people, especially those belonging to the BPL category, Mobile Fair Price Shops in tribal settlements, ensure quality of the food grains distributed through the TPDS, distribution of wheat flour in packed condition instead of wheat through the Fair Price Shops are recommended to address the shortcomings and weaknesses of the TPDS vis-avis the tribal population in Kerala.

Studies on food and feeding of marine demersal finfishes with special reference to trophic interactions

Food and feeding habits of fourteen demersal finfishes exploited off the Karnataka coast were studied to investigate trophic interactions within the marine food web. Index of Relative Importance (lRI),Ontogenetic, seasonal (pre-monsoon, monsoon and post-monsoon) variation in feeding and prey-predator relationship studies were conducted.The results of prey-predator trophic interaction studies identified four major trophic guilds based on the predators feeding similarity.Trophic guild I is 'copepod and detritus fceders'with an average group similarity of 61.4%. The second trophic guild, 'prawn and crab feeders'with an average similarity of 52.7%. 'Acetes feeders', the largest trophic guild with an average group similarity of 62.5%, composed of six demersal finfish species.The guild 'piscivores' is constituted by C. limba/us and P. arsius with an average similarity of 45%.For each predator, ontogenetic diet shift is common and is characterized by prey of low to high trophic level.Strong selection of certain prey types was observed in some predators while most of them avoided abundant prey.In addition to Acetes spp, strong predation impact was observed for penaeid prawns, epibenthic crabs and detritus.This information on trophic guilds and prey-predator interactions can be used to construct trophic model on the benthic ecosystem off Karnataka and to investigate fishery induced changes as well as predation impact of different animals on commercially important demersals

Studies on coagulase-positive staphylococci and Vibrio parahaemollticus in selected items of fish, crustaceans and fishery products

In India much work has been done on different aspects of quality of processed fish and fishery products (Pillai et al., 1965; Iyer and Chaudhuri, 1966; Iyer et al., 1966; Sreenivasan and Joseph, 1966; Pillai and Rao, 1969; Mathen et al., 1975; Valsan et al., 1985) and standards have been formulated for almost all the fishery products. Quality standards for fresh fish cover only a few fish species available in the markets. For the formulation and recommendation of quality standards for fish and fishery products for domestic trade detailed background informations are necessary. Khot et al., (1982), Valsan et al. J (1985) and Iyer et al. (1986) have reported on the bacterial flora of sea foods at retail level in Bombay. Cochin is a major fish landing centre of the south west coast of India, and an average of 25,000. tonnes of fish/shell fish are landed annually at this harbour which is about 10 percent of the total catch of fish in Kerala (Lakshmanan -et -al., 1984). All the varieties of marine, brackish water and fresh water fishes are available in Cochln throughout the year for catering to Its cosmopolitan. population and informations regarding their bacteriological quality are scanty. Hence the present study was undertaken to investigate the bacteriological quality of fish and fishery products available in the markets and cold storages situated in and around Cochin meant for internal consumption.

Thermal stability of myofibrillar protein from Indian major carps

The characteristics and stability of natural actomyosin (NAM) from rohu (Labeo rohita), catla (Catla catla) and mrigal (Cirrhinus mrigala) were investigated. The total extractable actomyosin (AM) was higher (7.60mgml−1) in the case of rohu compared with that from catla and mrigal (5mgml−1). Although the specific AM ATPase activity was similar (0.43–0.5 μmolPmin−1 mgP−1) among the three species, the total ATPase activity was lower in mrigal (25 μmol g−1 meat) compared with the other species (37 μmol g−1 meat). The inactivation rate constants (kd) of AM Ca ATPase activity showed differences in the stabilities of actomyosin among these fish, the actomyosin from catla being least stable. The NAM from these species was stable up to 20 ◦C at pH 7.0. Catla AM became unstable at 30 ◦C, while rohu and mrigal AM could withstand up to 45 ◦C. The thermal denaturation with respect to solubility, turbidity, ATPase activity, sulphhydryl group and surface hydrophobicity showed noticeable changes at around these temperatures

Rheological characteristics of suwari and kamaboko gels made of surimi from Indian major carps

The gel strength, compressibility and folding characteristic of suwari (set) and kamaboko (set and cooked) gels prepared from rohu (Labeo rohita), catla (Catla catla) and mrigal (Cirrhinus mrigala) surimi were examined to understand the occurrence of suwari and modori phenomena in surimi from major freshwater carps. Suwari setting of gels did not take place at lower temperatures. Suwari gels showed good gel strength at 50 C for rohu and at 60 C for catla and mrigal after 30 min setting time

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.

The Food Crisis: Implications for Labor

Food prices have gone up to prohibitive levels for many of the world’s poor. The vast majority of those who are hungry in the world today are working in agriculture, either as small landholders or as waged agricultural workers. The majority of the food producers have not benefited from rising prices. Apparently, the bargaining power of many producers, just as that of the end consumers, has been weakened vis-à-vis the buyers and retailers of agricultural produce. This powerlessness is also in the face of governments that fail to provide an appropriate infrastructure for smallholders and social protection. The first part of the book provides an introduction to the immediate and structural causes of the food crisis. The second part contains contributions that not only highlight the plight of rural labour but also develop tools for measuring the decent work deficit. The last part emphasizes income security as a major precondition for food security. It looks at the experiences of Brazil and India with the extension of social protection for the poor.

Perception of quality in certified organic pineapples by farmers in Kayunga district, Central Uganda: Implications for food security

In East Africa, Uganda is one of the major producers of organic pineapples for export. These pineapples are mainly produced in central Uganda and have to meet stringent quality standards before they can be allowed on international markets. These quality standards may put considerable strain on farmers and may not be wholly representative of their quality interpretation. The aim of this paper is therefore, to determine the Ugandan organic pineapple farmers’ quality perception, the activities they carry out in order to attain that quality and challenges (production, postharvest & marketing) faced on the same. Qualitative semi-structured interviews were carried out among 28 organic pineapple farmers in Kayunga district, central Uganda. Findings suggest that quality of organic pineapples is mainly perceived in terms of product attributes particularly appearance followed by food security provision. Certification plays a minor role in what farmers describe as organic quality. High production input costs (labour and coffee husks) coupled with a stagnant premium are some of the major challenges faced by farmers in attaining organic quality. The paper argues that currently there are concealed negative food security effects embroiled in these pineapple schemes. It is recommended that the National Organic Agricultural Movement of Uganda (NOGAMU) works with all relevant stakeholders to have the farmer premium price raised and an official organic policy enacted.

Development of molecular-based techniques for the detection, identification and quantification of food-borne pathogens

La presencia de microorganismos patógenos en alimentos es uno de los problemas esenciales en salud pública, y las enfermedades producidas por los mismos es una de las causas más importantes de enfermedad. Por tanto, la aplicación de controles microbiológicos dentro de los programas de aseguramiento de la calidad es una premisa para minimizar el riesgo de infección de los consumidores. Los métodos microbiológicos clásicos requieren, en general, el uso de pre-enriquecimientos no-selectivos, enriquecimientos selectivos, aislamiento en medios selectivos y la confirmación posterior usando pruebas basadas en la morfología, bioquímica y serología propias de cada uno de los microorganismos objeto de estudio. Por lo tanto, estos métodos son laboriosos, requieren un largo proceso para obtener resultados definitivos y, además, no siempre pueden realizarse. Para solucionar estos inconvenientes se han desarrollado diversas metodologías alternativas para la detección identificación y cuantificación de microorganismos patógenos de origen alimentario, entre las que destacan los métodos inmunológicos y moleculares. En esta última categoría, la técnica basada en la reacción en cadena de la polimerasa (PCR) se ha convertido en la técnica diagnóstica más popular en microbiología, y recientemente, la introducción de una mejora de ésta, la PCR a tiempo real, ha producido una segunda revolución en la metodología diagnóstica molecular, como pude observarse por el número creciente de publicaciones científicas y la aparición continua de nuevos kits comerciales. La PCR a tiempo real es una técnica altamente sensible -detección de hasta una molécula- que permite la cuantificación exacta de secuencias de ADN específicas de microorganismos patógenos de origen alimentario. Además, otras ventajas que favorecen su implantación potencial en laboratorios de análisis de alimentos son su rapidez, sencillez y el formato en tubo cerrado que puede evitar contaminaciones post-PCR y favorece la automatización y un alto rendimiento. En este trabajo se han desarrollado técnicas moleculares (PCR y NASBA) sensibles y fiables para la detección, identificación y cuantificación de bacterias patogénicas de origen alimentario (Listeria spp., Mycobacterium avium subsp. paratuberculosis y Salmonella spp.). En concreto, se han diseñado y optimizado métodos basados en la técnica de PCR a tiempo real para cada uno de estos agentes: L. monocytogenes, L. innocua, Listeria spp. M. avium subsp. paratuberculosis, y también se ha optimizado y evaluado en diferentes centros un método previamente desarrollado para Salmonella spp. Además, se ha diseñado y optimizado un método basado en la técnica NASBA para la detección específica de M. avium subsp. paratuberculosis. También se evaluó la aplicación potencial de la técnica NASBA para la detección específica de formas viables de este microorganismo. Todos los métodos presentaron una especificidad del 100 % con una sensibilidad adecuada para su aplicación potencial a muestras reales de alimentos. Además, se han desarrollado y evaluado procedimientos de preparación de las muestras en productos cárnicos, productos pesqueros, leche y agua. De esta manera se han desarrollado métodos basados en la PCR a tiempo real totalmente específicos y altamente sensibles para la determinación cuantitativa de L. monocytogenes en productos cárnicos y en salmón y productos derivados como el salmón ahumado y de M. avium subsp. paratuberculosis en muestras de agua y leche. Además este último método ha sido también aplicado para evaluar la presencia de este microorganismo en el intestino de pacientes con la enfermedad de Crohn's, a partir de biopsias obtenidas de colonoscopia de voluntarios afectados. En conclusión, este estudio presenta ensayos moleculares selectivos y sensibles para la detección de patógenos en alimentos (Listeria spp., Mycobacterium avium subsp. paratuberculosis) y para una rápida e inambigua identificación de Salmonella spp. La exactitud relativa de los ensayos ha sido excelente, si se comparan con los métodos microbiológicos de referencia y pueden serusados para la cuantificación de tanto ADN genómico como de suspensiones celulares. Por otro lado, la combinación con tratamientos de preamplificación ha resultado ser de gran eficiencia para el análisis de las bacterias objeto de estudio. Por tanto, pueden constituir una estrategia útil para la detección rápida y sensible de patógenos en alimentos y deberían ser una herramienta adicional al rango de herramientas diagnósticas disponibles para el estudio de patógenos de origen alimentario.

The potential for competitive and healthy food chains of benefit to the countryside

Agricultural policy liberalisation, concern about unhealthy diets and growing recognition of the importance of sustainable land use have fostered interest in the development of competitive food chains based around products that are beneficial to the rural environment. We review the potential for foods with enhanced health attributes based on alternative varieties/breeds and production systems to traditional agriculture which has been predominantly motivated by yields. We concentrate on soft fruit, which is an important source of polyphenols, and grazing livestock systems that have the potential for improving fatty acid profiles in meat products and find there to be clear scientific potential, but limited research to date. Consumer research suggests considerable acceptance of such products and willingness to pay sufficient to cover additional production costs. Purchase of such foods could have major implications for agricultural land use and the rural environment. There is little research to date on specific healthier food products, but spatially explicit models are being developed to assess land use and environmental implications of changing demand and husbandry methods.

Importance of mycotoxins in food and feed in India

Various food and feed samples including groundnut seed, maize, sorghum, soyabean cake, groundnut cake, cotton cake, poultry feed, buffalo milk, cow milk and milk powders were collected from farmers' fields, farmer's stores, oil millers storage, traders' storage, retail shops and supermarkets. More than 2000 samples were analysed by ELISA and most of the commodities, with the exception of sorghum seed, contained high levels of aflatoxin. Groundnut cake was one of the major cattle feed ingredients in the peri-urban area of Hyderabad (Andhra Pradesh, India) and >75% of the samples contained >100 µg/kg aflatoxin, leading to a high level of aflatoxin M1, in milk samples. Strategies to reduce aflatoxin levels (especially in groundnut) by management interventions at preharvest, harvest and storage, are discussed.

A framework for revitalisation of rural education and training systems in sub-Saharan Africa: Strengthening the human resource base for food security and sustainable livelihoods

Recent studies of the current state of rural education and training (RET) systems in sub-Saharan Africa have assessed their ability to provide for the learning needs essential for more knowledgeable and productive small-scale rural households. These are most necessary if the endemic causes of rural poverty (poor nutrition, lack of sustainable livelihoods, etc.) are to be overcome. A brief historical background and analysis of the major current constraints to improvement in the sector are discussed. Paramount among those factors leading to its present 'malaise' is the lack of a whole-systems perspective and the absence of any coherent policy framework in most countries. There is evidence of some recent innovations, both in the public sector and through the work of non-governmental organisations (NGOs), civil society organisations (CSOs) and other private bodies. These provide hope of a new sense of direction that could lead towards meaningful 'revitalisation' of the sector. A suggested framework offers 10 key steps which, it is argued, could largely be achieved with modest internal resources and very little external support, provided that the necessary leadership and managerial capacities are in place. (C) 2006 Elsevier Ltd. All rights reserved.