Véronique Bontemps

Soap Factories in Nablus: Palestinian Heritage (turâth) at the Local Level

Introduction

In an exhibit called Present Tense, the British-Palestinian artist Mona Hatum drew the map of the Oslo Accords on pieces of Nâbulsî soap, using small red pins.

More insidious than destructive acts or military invasions, the map is also a form of violence. It underlines here the very violence that emerged from the Oslo Accords, 1 that is, the fragmentation of the Palestinian Territories. The pieces of soap on which the map is drawn represent the essential but fragmented character of Palestinian identity. At the same time, while looking at this strange map, one can breathe the familiar smell of olive oil, and Nâbulsî soap is indeed famous for not melting so easily. White and cube-shaped, Nâbulsî soap stays and does not change. Soap, as a metaphor of heritage seems however, to be continuously threatened with dissolution or of being destroyed. The work of art thus discreetly underlines the continuous internal resistance (sumûd) that characterizes Palestinian society under occupation. Nâbulsî soap is therefore used as a metaphor to express a wider identity: that of the Palestinian people and their resilience on the ground, despite the material and symbolic violence they endure in everyday life. This paper discusses the various levels of Palestinian heritage (turâth) through the example of a particular item of Palestinian culture: soap and soap-factories in Nablus. It aims at understanding how this product of a small local industry has been and is still constructed as a material expression of heritage.

When thinking about Palestinian heritage, one immediately sees it within the context of a conflict with the Israeli occupier in terms of territorial legitimacy; yet, this political aspect obscures the fact that Palestinian heritage is itself multi-layered. In this paper, I argue that there is a dissonance between a narrative of national resistance to Israeli occupation, in which Nâbulsî soap has recently been included; and local perceptions and representations that show it as an aspect of local, familial and urban heritage. The coexistence between these two sets of discourses reflects the tension between "authorized discourse 2 " on heritage, as shaped by social and intellectual authorities in Palestine throughout the 20 th century on the one hand; and the particular relation to heritage, as constructed in everyday life by local individuals, on the other hand. But it also expresses, as I argue, the inherent "multivocacy" 3 of the Palestinian field of heritage through the various levels in which a particular item can be claimed as such: between affirmation of local anchorage, familial belonging and broader claims for national identity.

My argument is that the recent inscription of Nâbulsî soap into a narrative of Palestinian resistance (sumûd) comes to obscure its importance as an aspect of local, familial and urban heritage. Moreover, by fixing Nâbulsî soap-production within a "traditional" identity, it also obscures the issue of its modernization: on the local level, soap-industry is first and foremost an economic activity. But if Nâbulsî soap has never really "modernized", it is not only for economic reasons. It also has to do with local representations of soap and soap-industry, which illustrate a tension between a product of everyday life and a signifier of the pastthus, as I argue, as an "in-between heritage".

Recent evolutions in the uses and reshaping of soap-heritage show however, how the national and the local can be combined in patrimonializing Nâbulsî soap and soap-factories, and how subaltern or alternative management of heritage on the part of NGOs or local associations can succeed today in reshaping it, and giving it new meanings. While targeting a particular (i.e. Western) public, these initiatives also promote new conceptions of heritage, which give priority to "adaptive re-use" 4 rather than to mere conservation of the past. 5

1°) Nâbulsî Soap Industry and Heritage in the Palestinian Context

This section summarizes the development of soap-industry in Nablus. 6 Besides providing chronological markers, it also gives an idea about the material basis from which Nâbulsî soap factories were constructed as a symbol of a local power base for the urban Nâbulsî bourgeoisie, and of the changing forces which maintained the need for its continuation, from its "golden age" until its decline in the second part of the 20 th century.

About Nâbulsî Soap

Soap making is a very old tradition in the Middle East, as well as in all the Mediterranean area. It still has a domestic production in villages; women make soap at home using the remaining olive oil from the yearly harvest. It developed in Nablus as an urban industry, as the ruling families of the city invested in this field from the 18 th century onward. Following a renovation program of these factories during the second half of the 19 th century, soap making became a flourishing industry; with more than thirty soap factories, Nablus was exporting soap in all the Middle East, mostly to Egypt. 7 This success was mostly due to the fact that Nâbulsî soap was made of olive oil, the primary agricultural product of the Nablus district. Nablus is also located near the east bank of the Jordan River where qilî, 8 the second most important raw material for soap, grew abundantly. At the time, soap production was a profitable investment. The impressive soap-factory buildings interspersed throughout the old city of Nablus became symbols of wealth and prestige for their owners. 9 In the 1930s, Nâbulsî soap experienced its first important setback. 10 Two reasons are usually given for this. First, the lack of legal protection for the name "Nâbulsî" caused numerous cases of counterfeiting, especially in Egypt. In order to face this problem, the soap 4 Ibid., p. 129. factories constituted themselves as registered companies with brand names 11 to ensure the quality of the soap. The second reason was the imposition of taxes on the import of soap by Syria and Egypt. In addition, competition intensified with the Jewish mechanized industry that succeeded in obtaining tax exemptions from the British Mandate. 12 The Shemen factory for oil and soap, in particular, was established in 1922 near Haifa with a capital of 25 000 pounds (that is, almost as much as the 24 soap-factories in Nablus combined). 13 Power relation patterns between soap-factory owners and workers changed in the 1950s along with the introduction of what has been called "green soap". This soap is made from jift oil, in other words, from the remains of pressed olives and pits. It was used to clean floors and for laundry. In turn, this soap made of cheaper oil made it possible for former workers to start new soap factories and produce soap for themselves. The 20 th century also saw transformations in the raw materials used in soap production: the qilî was replaced by caustic soda in the 1920s and, even more importantly, olive oil is now being imported from Italy, mostly for economic reasons. The traditional process of production, however, has not changed significantly.

The first Intifada marked the final decline for soap-industry. Green soap factories of smaller size were the victims of the introduction of detergents and washing machines. For many of the old city"s factories, the work became dangerous as the old city became the target of Israeli attacks during the first Intifada. Thus, many of them closed in the 1990s. But the most important reason for this decline is probably the competition by cheaper foreign soap products (such as Lux and Palmolive), as well as the introduction of new consumption models that led young generations to prefer shampoo and perfumed soaps.

The situation has even worsened since the outbreak of the second Intifada in September 2000, and the reoccupation in 2002 of the major cities of the West Bank by the Israeli army. This time, however, there was renewed local interest for soap-factories. After they had been targetted by Israeli assaults, new discourses emerged that glorified Nâbulsî soap as a symbol of resistance to occupation, thus inscribing it in the broader narrative of Palestinian heritage (turâth) where it had not figured prominently until then.

In the next section, I give an overview of the "authorized discourses" 14 on heritage (turâth) in Palestine, according to the way that Palestinian folklorists have historically shaped them. This should in turn help us understand the meaning of this belated inclusion of Nâbulsî soap and soap-factories into a national narrative of Palestinian turâth.

Aspects of Palestinian Heritage: From the 'Nativist Ethnographers' of the British Mandate to the Struggle against Occupation

In her book about pilgrimages and nationalism in Palestine, 15 Emma Aubin-Boltanski 11 These brands are often symbols or names of animals: Muftahayn (the two keys), al-Jamal (the camel), al-Na"ama (the ostrich), al-Najma (the star), al-Baqara (the cow), al-Badr (the full moon), al-Asad (the lion) and others. Slogans were also added on the packaging such as "al-Sâbûn al-Nâbulsî al-Mumtâz" (Nablus soap extra-fine) or "al-Ma"rûf" (the famous one). describes the important place of folklore (fulklûr) in the Palestinian society. She also underlines the fundamental role of folklorists (these "Palestinian intellectuals and scholars, interested in heritage and local tradition" 16 ) in the process of constructing an object or a practice as being part of turâth. Palestinian folklorists took charge of the formation and shaping of a common national heritagesuch an action was all the more important in Palestine, where national identity was both denied and attacked by another national identity related to the Zionist undertaking.

Scholars generally consider that the beginning of a discourse on turâth (heritage) in Palestine is to be found in the "Nativist ethnography" 17 of Tawfiq Canaan and his circle 18 during the British Mandate. These Palestinian intellectuals from Jerusalem were writing mainly within the framework of learned societies, addressing the political elite of the Mandate as well as archaeologists and Western historians. They organized what is considered to be the first authorized discourse on turâth, through the idea that the Palestinian peasant represents a "living example" of everyday life as it was lived during Biblical times. 19 Convinced that the essence of Palestinian cultural heritage, as personified by its peasant heritage, was endangered by the penetration of Western modernity, they assigned themselves the task and responsibility to collect, classify and describe this already disappearing culture.

The works of these folklorists have been criticized (mostly by Palestinian scholars) for the "essentialism" and "reductionism" of their theses. Nevertheless, they produced a very rich ethnography, deeply sensitive to the "details of local practices, to variations over time (…) and to the interpretation of these practices in their wider social context" 20 .

These works are to be understood within the framework of what Tamari calls a "protonationalist" 21 context, at a key period (the British Mandate) when Palestinian modern identity was being forged. 22 Their works contain elements that would stay at the center of conceptions about Palestinian popular heritage until a very recent date: the idea of Palestinian identity as fundamentally related to the peasant. Even if the latter folklorist movements relied on an ideology proclaiming the existence of Canaanite roots (prior to Judaism) in the reconstruction of Palestinian identity, 23 the idea that the peasant is the authentic soul of the Palestinian nation 24 would stay as a key concept in the movements of national liberation in the 1970s and the 1980s.

After the nakba (disaster, catastrophe) 25 Canaan and his colleagues did not constitute a proper group but they shared a certain number of attributes, for example, they placed Biblical parallels and references at the core of their observations of the Palestinian popular culture (Salim Tamari, op. cit.). occupied the West Bank and the Gaza Strip, the ethnographic work of Canaan was revived by a folklorist movement intimately linked to the project of national liberation, cultural resistance to Israel and colonization. This movement aimed at collecting all the elements of "popular heritage" (al-turâth al-sha"bî) and folklore (fulklûr), including immaterial heritage such as oral literature, songs, folkloric dances, know-how and crafts (e.g. pottery, ceramics and embroidery). The infrastructure of the movement was formed by associations coming from the civil society, which were mostly women"s groups and research centres 26 that linked preservation of popular heritage to political emancipation. 27 This folklorist movement contributed toward nourishing the image of a Palestinian heritage fixed in a rural and peasant identity. In this respect, while the process of domestic soap-making by women in the village found its natural place in this context, 28 folklorists did not pay much attention to soap production in Nablus, an industry mostly run by the urban wealthy bourgeoisie. 29 The Nablus municipality published the first comprehensive study about Nâbulsî soap industry in 1999. 30 If its author, a Nâbulsî local historian, wrote it during the first Intifada (1987-1993), the date of publication is significant. The period that followed the Oslo Accords was indeed that of a renewed interest for heritage on the part of the new Palestinian Authority. For the new leaders of a proto-state, it was a means to assert its legitimacy for the purpose of bringing together and unifying diverse elements of a Palestinian national identity. 31 As Chiara Di Cesari puts it, heritage framed as a "national problem" was deeply intertwined with the broader project of building the Palestinian nation-state: 32 its peculiarities and varieties had to find a place in the large spectrum of resources which characterized Palestinian nationalism. Significantly, in the preface of the book, a former mayor of Nablus explains that the Nâbulsî soap industry has gained "an international reputation (…) as a craft linked to Palestinian oil", 33 thus explicitly giving a national dimension to Nâbulsî soap through this reference to Palestinian and not olive-oil.

Nâbulsî Soap Endangered: Soap-Factories as a Symbol of Sumûd

The outbreak of the Second Intifada and the reoccupation of the major cities of the West Bank by the Israeli army were a severe blow to the Nâbulsî soap industry. Besides the drop in exportations (from 1,200 to 700 or 800 tons per year), two old soap factories were totally destroyed in the old city of Nablus. This period saw the birth of new discourses on the soap factories, emphasizing the need to preserve their memory as visible traces of the past. Articles in the local newpapers and on internet started to refer to Nâbulsî soap as a vestige of a glorious past, an icon of Palestinian culture and national heritage, as well as the symbol of Palestinian resistance to occupation.

A Master dissertation defended at al-Najah University in Nablus on the architecture of the Nâbulsî soap factories asserts that "(…) there is an urgent need to gather information about soap factories before they are totally destroyed and this information lost forever (…)." 34 Later on, the author stresses the need to "preserve Palestinian heritage on the ground, in order to stand up to Israeli politics which try to replace this authentic identity, and to destroy its features". 35 In 2005, in the conclusion of an article entitled "Nâbulsî soap, ambassador of Palestinian heritage", the author evoked a wall of soap that was built by a Palestinian delegation in a congress at the Dead Sea. According to her, it was a way to "(…) show another aspect of this struggling people, and to give a good image of Palestinian heritage". 36 Here, Nâbulsî soap became Palestinian soap as a symbol of national and cultural identity, in struggle for Palestinian rights: in other words, an example of sumûd (steadfastness). This kind of rhetoric referring to the Palestinian heritage as being in danger of destruction is not new. Indeed, as mentioned previously, it was quite a common topic to the discourse on heritage and folklore in Palestine. What is relatively new, however, is the fact of placing soap factories into this heritage, by using rhetorical references of "urgency" and "rescue of culture". Nonetheless, this ideological discourse which fixes Nâbulsî soap heritage as a frozen symbol of resistance to occupation came to obscure its various meanings and significance on the local level, as well as its development through time. Before becoming a symbol, Nâbulsî soap was the product of the local industry, as well as a consumer product. It is therefore through these various meanings, as well as the way that the inhabitants of Nablus experience the development of soap as a symbol, that we can seek to understand how it is constructed as heritage through everyday practices and representations.

In the following section, various practices of Nâbulsî soap are presented. Both as a local industry and a heritage, it is facing a fundamental dilemma: namely, that it has to modernize to survive. Yet, if it modernized, as the inhabitants of Nablus say, "It would not be Nâbulsî soap anymore". By examining how ordinary people consider Nâbulsî soap in their everyday representations and practices, I try to show that this dilemma is constitutive of the meaning of Nâbulsî soap-industry as an "in-between" heritage.

2°) Practices of Nâbulsî Soap

When I started my fieldwork in Nablus in 2005, soap was still produced manually 37 in the three large factories 38 which were still operating: the Tûqân, the Masrî and the Shaka"a 39 factories. Around 80 % of Nâbulsî soap production is exported to Jordan, relying on old family ties between Nablus and the East Bank of the Jordan River. In Nablus, along with knâfa (a tasty pastry made of cheese and sugar), soap is still considered to be a specialty of the city. Indeed, it is still renowned in the Middle East for its quality.

In 2005, the soap industry was doing rather poorly. Supplying the factories with raw materials was getting increasingly difficult. Year after year, sales declined and profits fell. Abû Amjad, the accountant at the Tûqân factory, criticized the Palestinian Authority for not protecting this local industry, either by funding it or exempting it of taxes. He also criticized the lack of interest on the part of the owners themselves. He accused them of neglecting their industry: they did not bother to advertise or to open new markets 40 , nor did they try to modernize it.

The reason for this problem lays in the fact that soap-factory owners preserve their factory more for symbolic than economic reasons. Indeed, the primary meaning of Nâbulsî soapheritage is related to family honour: for important families in Nablus, soap factories are first and foremost linked to familial heritage, as a marker of their long-established urban identity.

'For the Name': Soap-Factories as a Family Heritage

As for Sabih al-Masrî, his politics on that matter is one of preservation (hifâz)… The first thing is that it belongs to our family"s heritage… And at the same time, it is the preservation of his father"s name. 41 Through the preservation of their factory, big families mean to preserve the name of their family: having a soap-factory is still a criterion of family greatness in Nablus. The soap factory is kept as a souvenir of this greatnes, and in recognition of the original activity of the family, which contributed towards building its wealth and social position. It is also a matter of social recognition and family honour within the framework of Nâbulsî urban society. As the accountant of the Masrî factory told me:

As I told you, it"s also that the soap factory has the name of the family (…) for the Masrî family to close its soap factory is shameful ("aîb), in the city… in Nablus… They will say: "Why is the Masrî family closing its factory? 42 Mahdi Ya"ish, a former soap producer whom I met in 2007, told me that the family factory was closed because of security reasons. Indeed, he feared that groups of young people (shabab) would enter the factory in the old city and use it as a base for armed operations. He told me: If it was not for the security situation, I would have it (the factory) re-opened. I would do it for the name. Because the idea is not to make money… I owe it to my grandparents, you know… This is what counts for me, I am not trying to make money. 43 The soap factory also has a symbolic meaning: it is a sign of historic urban belonging that is woven into the fabric of the old city, with all the familial prestige that this entails. To own a soap factory, one of the symbolic pillars of the Nâbulsî nobility until the twentieth century, is to distance oneself from the nouveau riche and newcomers in the city. Despite its decline, it is 40 They relied on the local Palestinian market which is gradually shrinking, and on long-existing exportation networks to Jordan. still an important sign of urban belonging because it is linked to the authenticity of the family name.

Today, however, the economic interests of the big families are of another dimension. For example, in 1994, the Hajj Tâhir al-Masrî company started to invest in several fields, along with the arrival of the Palestinian Authority. Soap factories are now only minor branches of the economic activity of the large families which own them. The paradox is indeed that while the preservation of heritage is one of the reasons for the survival of Nâbulsî soap factories, it is, at the same time, the reason for their decline. For the Tûqân, Shaka"a and Masrî families, the preservation of familial heritage relies on affective ties and a strong feeling of belonging to their city, but they are not willing to invest and modernize the soap factories, or to promote their heritage. They keep the factory open without trying to keep it alive.

"It did not Work": The Paradox of Modernization and 'Authenticity'

Why did the Nâbulsî soap industry never really modernize? There have actually been attempts throughout the 20 th century to mechanize the production and vary the ingredients, even though some factory owners sometimes seemed reluctant. As early as 1916, Rafîq Tamîmî and Mohammad Bahjat, two Ottoman officials who had been sent to collect information and write a guide about the southern half of the Beirut district, 44 wrote that the tools used for soap making in Nablus were "primitive". They added that the soap-factory owners should introduce proper tools and material in their industry. [START_REF] Bahjat | Rafîq Tammîmî, Wilâyat Bayrût, al-qism al-janûbî[END_REF] In 1927, Antonin Jaussen visited the Tûqân soap factory and made a similar comment. He saw the conservatism of the soap-factory owners as a common feature of the "Oriental mentality", and their "narrow-mindedness" as typically Nâbulsî. [START_REF] Jaussen | Naplouse et son district[END_REF] Later on, in an article on the political economy of the district of Nablus, Sarah Graham-Brown considered the soap industry in Nablus as a perfect illustration of "the effects of the changing structure within which the agriculturally based industries had to operate and of the inability or reluctance of their owners to adapt to it". [START_REF] Graham-Brown | The Political Economy of the Jabal Nablus, 1920-1948[END_REF] In 1923, Hajj Nimr al-Nâbulsî was the first soap-factory owner to import machines. Later on, in the 1950s and 1960s, producers from the Nâbulsî and the Kana"ân families imported machines from England and Germany. During my fieldwork, workers at the Masrî and Shaka"a soap factories showed me samples of soap to testify to the attempts made by the owners to mechanize the work and make soap "like Lux" 48but in the end, the conclusion was always the same: "It did not work".

It is indeed a widespread opinion in Nablus that "the problem of the Nâbulsî soap factories is that they did not renew their work". 49 "Renewal", "improvement, "development", "modernization"… a multiplicity of terms are employed, all converging to the same idea: should we look at the fabrication process or at the shape or packaging of Nâbulsî soap, the problem of its competitiveness is often attributed to its difficulty to face "modernity".

The inhabitants of Nablus themselves often attribute the lack of modernization of Nâbulsî soap to a problem of "mentality" ("aqliyya). As a member of the Shaka"a family once told me:

The problem with those people working with soap… they work with the old mentality (al-"aqliyya al-qadîma) of my father and grandfather… Some of them tried to think about how to develop… They tried, they didn"t succeed, they gave up. Amîn (director of the Tûqân soap factory), you tell him to change, he tells you no. He tells you it will not be Nâbulsî soap anymore. If you go and see Amîn"s client and tell him that Muftahayn soap 50 has become like that, he will tell you "No, it"s not Muftahayn soap, I don"t want it". 51 If Nâbulsî soap did not "develop", it is also because changing one or another of its characteristicsin the eyes of a customer who is used to it -would be to change its "identity": "it would not be Nâbulsî soap anymore". In order to better understand the difficulties of modernizing Nâbulsî soap, we have to open the black box of its "identity" and authenticity, by presenting some features of this identity that are both material and symbolic. This is not to suggest that Nâbulsî soap has an essential and authentic "identity" that should be discovered or unfolded. In fact, what makes Nâbulsî soap a form of heritage, and what is constitutive of it, is not so much the object in itself but rather the relation that people develop to it, their use of it, and the set of representations that they create about it.

In the next section, I present some local representations about what isor should be -Nâbulsî soap; that is to say, some collective images that define the idea of "heritage" for Nâbulsîs. For this I rely on local literature as well as on conversations and observations. Written sources often converge with memories of the inhabitants, creating images and representations that constitute a common base of references, which Maurice Halbwachs calls "social frames of memory". 52 Through these frames, which are constantly constituted and reconstituted by the community according to Halbwachs, individuals depict their signs and references in order to organize their memories. These collective frameworks engender representations without being explicitly present in the minds of the people. They often fix Nâbulsî soap as an "object of the past", which sometimes contradicts their practices of a product of everyday use.

Features and Representations: Nâbulsî Soap as an Object of the Past

History books, memoires and autobiographies on Nablus all have a passage on the soap industry, from Ihsan Nimr"s (the famous local historian) Tarîkh Jabal Nâblus wa al-Balqa" (History of Nablus and the Balqa") 53 to the autobiography of Muhammad Darwaza, Mi"at "âmm filastîniyya (One Hundred Palestinian Years) 54 to the Nâbulsî poet Mâlik Masrî"s Nâbulsiyyât (Things of Nablus) 55 .

When referring to Nâbulsî soap, these written sources generally evoke several aspects of the industry: the materials and ingredients formerly needed to make soap (that is to say, olive oil and qilî); the huge buildings of the soap factories; the soap itself with its basic cubic shape; and, finally, the manufacturing handmade process. Through these representations, the soap (as The reference to olive oil anchors Nâbulsî soap in a locality as well as in a sacred tradition, as olive trees and olive oil are mentioned in the Quran. 56 Most people in Nablus actually seem unaware of the fact that Nâbulsî soap is no longer made with local Palestinian olive oil. I noticed this when speaking at a conference at the French cultural center in Nablus in 2005: I explained that in the three large factories still operating in the city, soap was produced nowadays with refined olive oil imported from Italy. This information surprised the audience. This example shows the ambiguity of representation and its tenacity: for Nâbulsî inhabitants, it is indeed the pureness and quality of olive oil that gives soap its particular characteristics.

The second ingredient traditionally used for soap making, the qilî, was a key-element for exchange and trade between the Bedouin tribes and the merchants of Nablus. The evocation of the qilî is a pretext for highlighting these commercial relations at a time when Nablus was the capital of the entire region, and a crossroad for caravans. 57 Some images are frequently and regularly evoked, such as the camel caravans that used to bring olive oil in goatskin flasks into the soap factories until the 1960s.

When remembering the activity of the soap factories in the old city, inhabitants often evoke a reconstructed harmony of social relations, which are supposedly lost today: for example, the habit of some factory owners to freely distribute the remnants of the olive pits which were used for fuel. Through its traditional ingredients, one can see the reconstruction of the prosperity of Nâbulsî soap, which was linked to a "golden age" (al-fatra al-dhahâbiyya) of the city itself; prosperity attested by the regular journeys of peasants to the soap factory in town, in order to sell their oil. When referring to "their" soap, Nâbulsîs often recall or 56 Le Coran, sura XXIV "The Light", Paris, Albin Michel, 1990, XXIV 35, p. 376. 57 Ihsan Nimr, op. cit.; John L. Burckhardt, Travels in Syria and the Holy Land, London, J. Murray, 1822. reconstruct a past which is more than often idealized, and which strongly contrasts the current economic situation of their city. 58 For owners of a soap factory but also for ordinary Nâbulsîs, preservation of Nâbulsî soap is not the maintenance of a living activity, but rather the memory of a disappearing era.

To 'Understand' this Soap: Nâbulsî Soap as an 'In-Between' Heritage

The actual decline of the Nâbulsî soap industry is also a consequence of the diminution of local consumption: people who still buy and use it today are getting older. While they remain adamant in praising the good qualities of "their" soap, they constitute a population of mostly persons aged sixty years or more. Younger generations, in particular, prefer to use shampoo or other foreign products that appear more "modern" and of better quality. As Abû Hishâm, a small soap producer, told me once: "People don"t understand this soap anymore. And the more people who understand it are dying... the more our work is diminishing." 59 To "understand this soap" is to know its characteristics and to appreciate its quality, which is linked with the purity of olive oil. According to Nâbulsî inhabitants, Nâbulsî soap is very efficient against dandruff, which makes it an excellent product to wash one"s hair. Another characteristic is that it does not foam easily but indeed, only when the skin is perfectly clean. 60 The materiality of this "identity" itself has a symbolic side: the issue here is not to question whether these qualities of Nâbulsî soap are real, but rather to stress the fact that older Nâbulsî inhabitants really believe in them, thus defining this "system of regulated dispositions" that Pierre Bourdieu calls a habitus. 61 An old Nâbulsî housewife polishes her pots with Nâbulsî soap -not because it"s nâbulsî but, as she will tell you, because the olive oil gives it a good shine. At the same time, by doing this, she is performing the socially recognized habitus of a Nâbulsî woman.

The relationship of Nâbulsîs to their soap is therefore deeply ambiguous. Many of them do not use it anymore. And for those who use it, Nâbulsî soap is far from being an anonymous product that can easily be transformed. It has a deeply affective dimension. For its customers, who are often older Nâbulsîs used to the soap"s shape and aspect, it expresses the memory of a bygone past, which keeps on surviving through the use of this soap. It is not a mere thing, but it crystallizes ties and time. Considered as archaic or unique, it can also become a sign of nostalgia or of remembrances of the past, "when Nablus was Nablus" as Nâbulsî inhabitants like to say.

Krysztof Pomian considers that an object (a "cultural good") is established as heritage according to a sequence: first, the object is a simple "thing" (a good to be used, i.e. a consumer good); it then becomes a "waste material" when it loses this function (as something to be used); it finally becomes a "semiophor" which means that "it bears visible characteristics capable of receiving new meanings". 62 It acquires a new function, which is no longer the utilitarian function of simple "things", but also a function to recall the past. Today, soap industry in Nablus is in an "in-between" situation: between preservation of its "traditional" identity and the necessity to survive and adapt to a globalized market. Its decline is partly due to the fact that it did not modernize. But if it changed to better fit the more "modern" requirements of everyday use, it would cease to exist as Nâbulsî soap: "It would not be Nâbulsî soap anymore". Nâbulsî soap thus occupies a liminal space between an everyday object and a heritage symbol: while it is still used as a thing in the present, it is also seen as a symbol of a glorious past. Its transformation into heritage is still an ongoing and slow process, changing little by little, without a central authority that would take this transformation in charge.

3°) Reinventing Nâbulsî Soap 1. A 'New Palestinian Past'

In the absence of real investment on the part of the Palestinian Authority or even by the owners themselves, independent structures and individual initiatives tried to take over this transformation of Nâbulsî soap as an element of heritage. For the past few years, there have been initiatives made by small groups, private foundations and local NGOs, which have decided to invest in Nâbulsî soap and soap factories.

These various initiatives have something in common: they try to escape the dilemma between preservation and modernization, by following new conceptions of heritage that emerged in Palestine in the 1990s, with the work of NGOs which focused primarily on the architectural heritage of the old urban centers of Palestine. This new generation of architects tried to restore and preserve the important architectural heritage of Palestine both from the erosion of time, and from the destruction caused by the occupation.

This field, which Di Cesari calls a "new Palestinian past", 63 focuses on a close and less essentialized past and on the re-use for the present rather than cautious conservation of the past. Associations such as the Centre for Cultural Heritage Preservation (CCHP) in Bethlehem and the Hebron Rehabilitation Committee (HRC) promote notions, at the core of their projects, like "adaptation for public use" or "adaptive re-use", which allow for the reshaping of heritage according to the needs of present time. 64 In Nablus, descendants of the famous soapmanufacturer Sheikh "Amr "Arafât created a small foundation in order to refurbish the old family soap factory. The architect Nâsir "Arafat (the grand-nephew of Sheikh "Amr) began to restore it in 2005 in order to create a small cultural center for children, with a small space dedicated to a soap museum. The aim of such an initiative is to re-use the factory buildings by adapting them to other practices and activities.

Back to the Authentic?

All these initiatives (associations and NGOs) recognize the "traditional product" not as the reproduction of a so-called heritage of the past, but rather as a social construction. They in turn share an assessment: they notice that the only major transformation recently introduced into the soap industry, namely, the use of industrial European oils instead of Palestinian olive oil, led to it being pulled out from its local roots. In order to revive the industry, it has to be re-anchored with its old roots. 63 "(…) a close past, (…) vernacular, lived, small-scale, centred on the home rather than the palace. Rather than displayed in a glass case, this is a past to be creatively put to use for socio-economic development, or, in other words, a site of cultural production for the present » Chiara Di Cesari, op. cit., p. 65. 64 See, for example, the site of CCHP: www.cchp.ps.

For these groups, the issue then is less to preserve Nâbulsî soap than to revive tradition by "returning" to the use of Palestinian olive oil. At the same time, they try to mechanize part of the process, in order to give soap a nicer and more attractive shape and aspect. In their opinion, returning (i"âda) to the roots of the industry, along with improving the shape of the soap in order to meet the demand of a "modern" public, could result in a successful "revival of tradition through innovation". 65 For Hâjj Mu"taz al-Nâbulsî, grand-nephew of the famous soap-maker Hâjj Nimr al-Nâbulsî (the first soap-maker to import machines), mechanization of the process linked to the re-use of local olive oil was the key to a possible "rebirth" of Nâbulsî soap. Hâjj Mu"taz al-Nâbulsî experimented with a different way to produce soap:

(…) with a shape and weight (100 grams) that would suit the local and foreign consumer, which would look like the Lux soap, and yet would be 100 % olive oil, and would have all the advantages of the Nâbulsî soap". 66 In order to do that, a "radical change" (as he said) was needed in the production process, because he did not want to introduce any material that would not be olive oil.

During the period of my fieldwork in Nablus, I followed the development of his attempts to "improve" Nâbulsî soap. He never showed me what he called his "laboratory" (where he made his experiments) but he gave me samples of his new soap, as well as a bottle of liquid soap (as he also did to any tourist passing by). Hâjj Mu"taz distributed his products in some drugstores, but the price was too high to allow for a proper diffusion. Photo 3: "Improved" Nâbulsî soap

A Solution through Export?

One small soap-manufacturer in Nablus told me once: "The Nâbulsî client wants something cheap, or he wants his piece of Nâbulsî soap." 67 Indeed, there seems to be no real market for this kind of "improved" soap in Nablus: in fact, all associations and NGOs involved in reshaping Nâbulsî soap turn to exportation. The public targeted by this reinvented product is, above all, a Western public: the idea is to take advantage of the current interest in natural products. In Syria, it is these new export networks toward the West that have allowed the revival and boom of the Aleppo soap industry during the past twenty years. Unlike those exported by the soap factories of Aleppo, however, the quantities of Nâbulsî soap exports are quite limited: two or three tons maximum, ordered by associations in France, Canada and Great Britain which support Palestinians.

In the extremely politicized and controversial context of Palestine, a significant amount (if not all) of the exports go through activist networks, which already import other products like olive oil. The idea is to produce a soap whose shape, aspect and packaging please the Western consumer while, at the same time, let him feel that he is buying a little piece of Palestine. Reference to olive oil, the major ingredient in the soap, enhances here both its traditional and national character. It engenders representations to a large extent stereotyped for some groups of activists, who have a certain idea of Palestine as "authentic" versus an aggressive "modernity" represented by Israel.

By re-inscribing Nâbulsî soap in its very local roots (the re-use of Palestinian local olive oil), these initiatives also promote it, for a Western public, to a national character. It is thus reembedded within the larger frame of sumûd, i.e. daily Palestinian resistance to Israeli occupation. Whereas Nâbulsî soap and the soap industry, as a representation of fixed heritage, refer first and foremost to the nostalgic memory of the city, it can be reinvested with new meanings that show the malleability and "multivocacy" of Palestinian heritage.

Conclusion

For heritage studies, the Palestinian field often appears like a laboratory in which to observe practices and discourses of heritage in the absence of a proper nation-state. But if there is no Palestinian state, there is nevertheless an authorized discourse on heritage, which was first defined by folklorists and then by the Palestinian National Authority. This authorized discourse is itself far from being monolithic: it is historically defined, more than often in reaction against the Zionist image, and then in relation to the Israeli occupier and the national struggle for a Palestinian state. There are also local, sometimes subaltern practices that appear in opposition to the remolding of this heritage, or simply in dissonance with the authorized discourse that defines the "true" objects of heritage. 68 Nâbulsî soap is indeed a very local heritage that people cherish because they are used to it, because they think olive oil is good for their skin, and because it reminds them of old and better times. It is difficult, however, to totally separate heritage from its political aspects. By examining local representations and practices, I tried to show how ordinary objects can take an officially inscribed national significance, while retaining other values and meanings in daily use, as well as the ways that everyday actions (such as scrubbing the dishes or washing one"s hair) can be infused with overtones of national heritage.

In the last part of this article, I also showed how, in the overwhelming context of territorial conflict, Nâbulsî soap is detoured away from its local meaning in order to be turned "towards a Western public" as an image and symbol of Palestine. This is another example of dissonance, but which shows that heritage is far from being a fixed object, and that "revival" or rebirth of heritage often means reshaping and creating by targeting a particular public.

68

For the examples of pilgrimages, see Aubin-Boltanski 2007.

  of 1948, but mostly after 1967 when Israel 16 Ibid., p. 161. 17 Salim Tamari, "Lepers, Lunatics and Saints: The Nativist Ethnography of Tawfiq Canaan and his Jerusalem Circle", Jerusalem Quartely, Vol. 5, no. 1, 2004, p. 25. 18 Tawfiq Canaan (1882-1964) was a doctor and the son of a Protestant Lutheran pastor. He published a great number of articles and books (both in English and in German) on Palestinian popular culture. During the 1920s and 1930s, his works, as well as those of his colleagues such as Khalil Totah, Omar Saleh al-Barghouti, Stephan Hanna Stephan and Elias Haddad) were published in the Journal of the Palestine Oriental Society.

19

  Emma Aubin-Boltanski, op. cit.; Chiara Di Cesari, op. cit. ; Salim Tamari, op. cit. 20 Salim Tamari, op. cit., p. 39. He in fact adds that the richness of the analysis often contradicts the essentialism of the concepts. 21 Ibid., p. 38. 22 Rashid Khalidi, Palestinian Identity: The Construction of Modern National Consciousness, Columbia University Press, 1997. 23 Salim Tamari, op. cit., p. 29. This Canaanite ideology is still vivid nowadays through certain festivals. It has nevertheless been criticized by Palestinian intellectuals such as Zakariyya Mohammad. 24 Nadine Picaudou (ed.), Territoires palestiniens de mémoire, Paris, Karthala, 2006 ; Ted Swedenburg, "The Palestinian Peasant as National Signifier", Anthropological Quarterly, 63 (1), 1990, p. 18-30. 25 For Arabs, this name refers to the creation of the state of Israel and the exodus of hundreds of thousands of Palestinian from their land, mostly to the surrounding Arab countries.

26

  The most famous amongst them is In"âsh al-Usra (Revival of the Family), founded by Samiha Khalil at the end of the 1960s. 27 Emma Aubin-Boltanski, op. cit.; Maher Charif, Le patrimoine culturel palestinien, Paris, Le Sycomore, 1980; Chiara Di Cesari, op. cit. 28 In 1973, the Centre of Research on Palestinian Heritage and Society in al-Bireh conducted and published a study on popular heritage in Palestine through the example of a specific village in the West Bank, Turmus"aya. The eleventh chapter in this study entitled: "Popular industry in the village" presents the soapindustry among other villager industries. The first issue of al-Turâth wa al-Mujtamaʽ ("Heritage and Society"), a journal published by the same centre, contains an article on "agricultural activities of the Birzeit peasant"; this article cites soap, which was mostly made by women, among the agricultural productions. 29 There is very little mention in the works of the folklorists in the 70s and 80s of Nâbulsî soap factories, except for an article in 1975 in al-Turâth wa al-Mujtama". Interestingly, the Encyclopaedia of Palestinian folklore (edited by the famous Palestinian folklorist Nimr Sirha) does not include a single notice on Nâbulsî soap, and only shows a single photograph of the pouring process (the bast) of soap in one of Nablus"s factories. 30 Husam Sharif, op. cit. 31 See Emma Aubin-Boltanski, op. cit. 32 Chiara Di Cesari, op. cit., p. 108. 33 Husam Sharif, op. cit., preface.

34

  Mona Salameh, "Amârat al-masâbin wa "alâqatuhâ bi-l-takhtît al-"imrânî li madînatNâblus al-qadîma (Soap Factory Buildings and their Relation to the Urban Fabric in the Old City of Nablus), Master in Architecture, al-Najâh University, Nablus, 2003, p. I (my translation). 35 Ibid., Introduction (my translation). 36 Mirvat "Aûf, Sâbûn Nâblus, safîr al-turâth al-filastînî (Nablus Soap, Ambassador of Palestinian Heritage), www.islamonline.net/servlet/Satellite?c=ArticleA_C&cid=1178724155783&pagename=Zone-Arabic-ArtCulture%2FACALayout (2 October 2009), article from the 22 September 2005 (my translation).37Except for some minor transformations. See Véronique Bontemps, op. cit. 38 Some small factories also functioned occasionally.39The owners are three large and wealthy families of Nablus.

41

  Interview with a member of Masrî family, Nablus, 2005 (my translation). 42 Interview with the accountant of the Masrî factory, Nablus, 2005, (my translation). 43 Interview with Mahdi Ya"ish, Nablus, 2007, (my translation).

50

  Photo 1: A soap-factory in the old city of Nablus

58

  Even if this situation seems to be a bit better today due to the opening, in July 2009, of the main checkpoints closing the city. 59 Interview with Abû Hishâm in Nablus, 2007 (my translation). 60 People often refer to a joke made by an Egyptian actor invited to Nablus in the 1960s by a member of the Shaka"a family. When offered a piece of Nâbulsî soap, he said: « So you have to wash with another soap first, and then use Nâbulsî soap! » 61 Pierre Bourdieu, Esquisse d"une théorie de la pratique, Paris, Seuil, 2000 (1 st ed. 1972). 62 Krysztof Pomian, « Musée et patrimoine », in Henri-Pierre Jeudy (ed.), Patrimoines en folie, Editions de la Maison des Sciences de l"Homme, 1990, p. 177-198.

65

  For several examples of this process in France, see Christian Bromberger & Denis Chevallier (ed.), Carrières d"objets. Innovations et relances, Paris, Editions de la Maison des Sciences de l"Homme, 1999. 66 Interview with Hâjj Mu"taz al-Nâbulsî, Nablus 2005 (my translation). 67 Interview with Mustafâ Tbeîla, Nablus 2006 (my translation).

5

  This paper relies on the fieldwork which I conducted in Nablus for my PhD between 2004 and 2008. I would like to thank PRIO-Cyprus for inviting me to present my research at its annual conference in Nicosia in 2010, as well as Roger Heacock who advised me to submit a paper for this conference.

	6	For a more extensive historical overview, see Beshara Doumani, Rediscovering Palestine. Merchants
	and Peasants in Jabal Nablus (1700-1900), London, University of California Press, 1995, and Véronique
	Bontemps,	Soap	from	Nablus.	A	small	history	of	Nâbulsî	soap,
	http://www.nablusguide.com/index.php?option=com_content&view=category&layout=blog&id=42&Itemid=65
	&lang=en.

7

Beshara Doumani, op. cit.

8

The qilî was obtained by burning a wild plant of the steppe and reducing it to ashes. 9 Beshara Doumani, op. cit. 10 Ali Nusuh Tâher, Shajarat al-zaytûn. Tarîkhuhâ, amrâduhâ, sinâʽatuhâ (The Olive Tree. History, Diseases, Industry), Jaffa, 1947; Husam Sharif, Sinâʽat al-sâbûn al-nâbulsî (Nâbulsî soap-industry), Nablus, Municipality of Nablus, 1999.


Helena A Soini

Donald Wiesler

Sachiko Koyama

Christophe Féron

Claude Baudoin

Milos V Novotny
email: novotny@indiana.edu

COMPARISON OF URINARY SCENTS OF TWO RELATED MOUSE SPECIES, Mus spicilegus and Mus domesticus

Keywords: Mus spicilegus, Mus domesticus, urinary volatile profile, gas chromatography/mass spectrometry, stir bar extraction, pheromones

Abstract -Whereas the house mouse (Mus domesticus) has been studied extensively in terms of physiology/behavior and their pheromonal attributes, the evolutionary related mound-building mouse (Mus spicilegus) has received attention only recently due to its divergent behavioral traits related to olfaction To this date, no hemical studies on urinary volatile compounds were performed on M. spicilegus. The rationale for our investigations has been to determine any potential differences in urinary volatiles of intact and castrated M. spicilegus male and explore further whether this species could utilize the same or structurally similar pheromones as the male house mouse, Mus domesticus. The use of capillary gas chromatography/mass spectrometry (GC-MS) together with sorptive stir bar extraction sampling enabled such quantitative comparisons between the intact and castrated M. spicilegus urinary profiles. Additionally, through GC-MS and atomic emission (sulfurselective) detection, we were able to identify distinct qualitative molecular differences between intact M. spicilegus and M. domesticus. On one hand, a series of volatile and odoriferous lactones and the presence of coumarin were the unique features of M. spicilegus, as was the notable absence of 2-sec-butyl-4,5-dihydrothiazole (a prominent M. domesticus male pheromone) and other sulfur-containing compounds. Castration of M. spicilegus males eliminated several substances, including -hexalactone and -octalactone, and decreased substantially additional compounds, suggesting their possible role in chemical communication. On the other hand, some other M. domesticus pheromones were also found in M. spicilegus urine. These comparative chemical analyses support the notion of metabolic similarities as well as the uniqueness of some volatiles for M. spicilegus, which may have a distinct physiological function in reproduction and behavior.

INTRODUCTION

There is an evolutionary connection between the common house mouse (Mus domesticus) and the mound-building mouse (Mus spicilegus) species, with two pairs of close species and subspecies in the phylogenetic tree, M. musculus domesticus and M. m. musculus in one pair and M. spicilegus and M. macedonicus in the second one [START_REF] Bonhomme | Biochemical diversity and evolution in the genus Mus[END_REF][START_REF] Sage | House mouse as models in systematic biology[END_REF]. The M. domesticus living environment is interwoven to human habitats, whereas M. spicilegus species is feral, and in some cases, at least during the summer period, M. spicilegus may live in a close contact with M. m. musculus [START_REF] Orsini | Le complexe d'espèces du genre Mus en Europe Centrale et Orientale. II. Critères d'identification, répartition et caractéristiques écologiques[END_REF][START_REF] Sokolov | Mus spicilegus[END_REF][START_REF] Simeonovska-Nikolova | Spatial organization of the mound-building mouse Mus spicilegus in the region of northern Bulgaria[END_REF]. Additionally, in their social structure and behavior, the M. spicilegus have developed far apart from the M. domesticus species. Unlike M. domesticus, M. spicilegus are monogamous [START_REF] Patris | Female sexual preferences differ in Mus spicilegus and Mus musculus domesticus: the role of familiarization and sexual experience[END_REF][START_REF] Dobson | Experimental tests of spatial association and kinship in monogamous mice (Mus spicilegus) and polygynous mice (Mus musculus domesticus)[END_REF][START_REF] Baudoin | Male-female associations and female olfactory neurogenesis with pair bonding in Mus spicilegus[END_REF][START_REF] Gouat | Deficit in reproduction in polygynously mated females of the monogamous mound-building mouse Mus spicilegus[END_REF] and build co-operatively colonial mounds for overwintering [START_REF] Orsini | Le complexe d'espèces du genre Mus en Europe Centrale et Orientale. II. Critères d'identification, répartition et caractéristiques écologiques[END_REF][START_REF] Garza | Social structure of the mound-building mouse Mus spicilegus revealed by genetic analysis with microsatellites[END_REF][START_REF] Poteaux | Socio-genetic structure of mound-building mice, Mus spicilegus, in autumn and early spring[END_REF]. Moreover, M. spicilegus males display intense paternal care [START_REF] Patris | A comparative study of parental care between two rodent species: implications for the mating system of the mound-building mouse Mus spicilegus[END_REF][START_REF] Féron | Paternal care in the mound-building mouse reduces inter-litter intervals[END_REF].

The M. spicilegus species appear to rely on olfaction in their social behavior and mate selection [START_REF] Patris | Female sexual preferences differ in Mus spicilegus and Mus musculus domesticus: the role of familiarization and sexual experience[END_REF][START_REF] Patris | Agonistic and sociable behaviors in the mound-building mice, Mus spicilegus: a comparative study with Mus musculus domesticus[END_REF]Heth et al., 2002[START_REF] Heth | Genetic relatedness assessment through individual odour similarities in mice[END_REF]. In this species, social and kin recognition through olfactory cues have been observed in several studies [START_REF] Gouat | Conspecific and heterospecific behavioural discrimination of individual odours by mound-building mice[END_REF][START_REF] Dobson | Experimental tests of spatial association and kinship in monogamous mice (Mus spicilegus) and polygynous mice (Mus musculus domesticus)[END_REF][START_REF] Baudoin | Male-female associations and female olfactory neurogenesis with pair bonding in Mus spicilegus[END_REF][START_REF] Busquet | Odour similarities as a basis for discriminating degrees of kinship in rodents: evidence from Mus spicilegus[END_REF][START_REF] Todrank | Preferences of newborn mice for odours indicating closer genetic relatedness: is experience necessary?[END_REF][START_REF] Colombelli-Negrel | Male and female mound-building mice, Mus spicilegus, discriminate dietary and individual odours of conspecifics[END_REF]. The major mammalian scent sources for chemical communication are sex glands, other skin glands or urine. The salivary androgen-binding proteins [START_REF] Laukaitis | Salivary androgen-binding protein (ABP) mediates assortative mate selection in Mus musculus[END_REF][START_REF] Talley | Female preference for male saliva: implications for sexual isolation of Mus musculus subspecies[END_REF] and tear fluid peptides [START_REF] Kimoto | Sex-specific peptides from exocrine glands stimulate mouse vomeronasal sensory neurons[END_REF] have also been reported as sources of chemical messangers in for M. domesticus. In the house mouse, the urine-mediated chemical signals have been relatively well-characterized over the years (for a review, see [START_REF] Novotny | Pheromones, binding proteins and receptor responses in rodents[END_REF][START_REF] Hurst | Scent wars: the chemobiology of competitive signaling in mice[END_REF]. To date, the chemical constituents of various scent sources for the M. spicilegus species, used in communication, are still unknown. Behavioral tests have revealed that the odor cues (expected to originate mainly from urine) are used in communication [START_REF] Patris | Female sexual preferences differ in Mus spicilegus and Mus musculus domesticus: the role of familiarization and sexual experience[END_REF][START_REF] Féron | Social regulation of reproduction in the female moundbuilder mouse (Mus spicilegus)[END_REF][START_REF] Busquet | Odour similarities as a basis for discriminating degrees of kinship in rodents: evidence from Mus spicilegus[END_REF][START_REF] Colombelli-Negrel | Male and female mound-building mice, Mus spicilegus, discriminate dietary and individual odours of conspecifics[END_REF]. Thus far, there have been no complementary reports on the chemical nature of such olfactory cues.

The first purpose of this study has been to structurally characterize and quantify individual chemical constituents of the urinary volatile profiles for the intact male M. spicilegus against those of castrated animals. The second goal was to compare the findings qualitatively to the previously well characterized male M. domesticus urinary compounds.

A quantitative comparison of the intact and castrated M. spicilegus male volatile profiles was performed to explore the metabolic end-products which could be produced under the testosterone control. Some of the endocrinologically controlled urinary constituents may act as chemical communication means for various reproductive and social functions.

Additionally, the urinary volatile profiles of female and male M. spicilegus individuals were qualitatively compared in order to explore the gender roles related to the urine-mediated chemical communication, such as those used in mate selection and individual recognition. This chemical characterization is expected to provide some clues to the observed differences in social and behavioral characteristics between the two species.

To facilitate this study, we used the stir bar aqueous extraction method [START_REF] Baltussen | Stir bar sorptive extraction (SBSE), a novel extraction technique for aqueous sample: Theory and principles[END_REF][START_REF] Baltussen | Sorptive sample preparationa review[END_REF], which was followed by solventless sample introduction into a gas chromatograph-mass spectrometer (GC-MS) instrument. This methodology is compatible with screening for volatile organic compounds at low concentrations in biological samples and is well suited for a compound identification and quantitative comparisons [START_REF] Soini | Stir bar sorptive extraction: a new quantitative and comprehensive sampling technique for determination of chemical signal profiles from biological media[END_REF]. In addition, a combination of gas chromatography with atomic emission detection (GC-AED) was utilized for the highly sensitive sulfur compound profiling. In this report, we also take our previously determined characteristics of the male M. domesticus (ICR, C57Black/B6 and C57Black/B10) urinary volatile components and compare them qualitatively with the new M. spicilegus chemical information obtained in this study.

Analytical approaches for the M. domesticus and M. spicilegus samples were identical. All analyses were performed in the same laboratory.

METHODS AND MATERIALS

Experimental Animals Mus spicilegus mice were fifth-generation animals from a population collected in Gyöngyös, Hungary, in October 1999. From the time of their collection, the genealogy of every individual was known and all breeding pairs had been formed in a way to avoid inbreeding. They were bred at the University of Paris 13 under laboratory conditions (20±1•C) with a 14 : 10 h light : dark cycle. Food (mouse pellets type M20, Special Diet Services, Witham, Essex, U.K.), water and bedding material (sawdust and cotton) were provided. Mice were weaned at 28 days of age and housed in same-sex sibling groups from 35 days of age. Males and females were 3-6 months old when they were used as urine donors.

For the male-female mouse comparisons, 6 males and 6 females were isolated in standard polycarbonate cages (26 x 16 cm and 14 cm high) 1 week before urine collection in order to eliminate the social dominance effect (Féron andBaudoin, 1993, 1998). With the male presence being required to induce sexual receptivity in M. spicilegus females [START_REF] Féron | Social regulation of reproduction in the female moundbuilder mouse (Mus spicilegus)[END_REF], we assumed that all the isolated females were in anoestrus.

For the comparisons between intact and castrated males, isolation occurred 3 weeks prior to urine collection. Twenty males were isolated in standard polycarbonate cages (26 x 16 cm, 14 cm high). Six of them were then castrated under anesthesia induced by an intraperitoneal injection of a mixture of ketamine (100 mg/kg per mouse) and xylazine (5 mg/kg per mouse).

Male M. domesticus urinary volatile profile from inbred ICR albino mice, C57BL /B6 and C57BL /B10 black mice (Jackson Laboratories, Bar Harbor, ME) were used for qualitative comparisons. The data obtained for M. domesticus have been previously reported by [START_REF] Harvey | Pattern of volatile compounds in dominant and subordinate male mouse urine[END_REF][START_REF] Novotny | Chemical identification of MHC-influenced volatile compounds in mouse urine. I: Quantitative proportions of major chemosignals[END_REF] Urine Collection Animals were introduced individually into a clean polycarbonate cage and surveyed for urination during 5 minutes at maximum. Fresh individual urine was then quickly collected with a syringe and frozen (-20°C). A preliminary study showed that this was the best process fitted to these very sensitive mice. Reagents and Analytical Methods All compound identifications were verified through the authentic standards whenever available. Standard compounds were either purchased from Aldrich Chemical Company (Milwaukee, WI) or synthesized in our laboratory according to the previously described synthetic methods for dehydro-exo-brevicomin (Wiesler et al., 1984), sec-butyl-4,5-dihydrothiazole [START_REF] North | Synthetic studies towards cyclic peptides. Concise synthesis of thiazoline and thiazole containing amino acids[END_REF]) and 6-hydroxy-6-methyl-3heptanone [START_REF] Novotny | A unique urinary constituent, 6-hydroxy-6-methyl-3-heptanone, has the puberty accelerating pheromone activity in mice[END_REF]. Twister™ stir bars (10 mm in length, 0.5 mm film thickness, 24-μL polydimethylsiloxane (PDMS) volume) were used as the sorptive extraction devices. They were purchased from Gerstel GmbH (Mülheim an der Ruhr, Germany).

Ethical Note

Volatile and semivolatile compounds were extracted from 0.2 ml of urine in 20-ml capped glass vials for 60 min with a Twister™ stir bar. The urine samples were first diluted with 2.0 ml water (high-purity OmniSolv water, EM Science, Gibbstown, NJ). As an internal standard, 8 ng of 7-tridecanone (Aldrich, Milwaukee, WI) was added in 10 l of ethanol to each vial. Stirring speed was 800+ rpm on the Variomag Multipoint HP 15 stirplate (H+P Labortechnic, Oberschleissheim, Germany). Prior to extraction, all glassware was washed with acetone and dried at 80° C. After extraction, the stir bars were rinsed with a small amount of distilled water, dried gently on a paper tissue, and placed in the TDSA autosampler tube (a product of Gerstel GmbH, Mülheim an der Ruhr, Germany) for the gaschromatographic (GC) or GC-MS analysis. GC equipment for the sulfur compound analysis consisted of an Agilent GC Model 6890 instrument with an Atomic Emission Detector (AED) (Model G2350A from Agilent Technologies, Wilmington, DE) and a Thermal Desorption Autosampler (TDSA, Gerstel). The separation capillary was HP-5MS (30 m x 0.25 mm, i.d., 0.25 μm film thickness) from Agilent. Samples were thermally desorbed in a TDSA automated system, followed by injection into the column with a cooled injection assembly, CIS-4. TDSA operated in a splitless mode. Temperature program for desorption was 20° C (0.5 min), then 60° C/min to 280° C (10 min). Temperature of the transfer line was set at 280° C. CIS was cooled with liquid nitrogen to -60° C. After desorption and cryotrapping, CIS was heated at 12° C/s to 280° C with the hold time of 10 min. CIS inlet was operated in the solvent-vent mode, a vent pressure of 14 psi, a vent flow of 30 ml/min, and a purge flow of 50 ml/min. The temperature program in the GC operation was 40° C for 5 min, then increasing to 200° C at the rate of 2° C/min. The final temperature was held for 10 min. The carrier gas head pressure was 14 psi (flow rate, 1.2 ml/min). The GC unit was operated in the constant-flow mode. The emission lines for carbon (193 nm), sulfur (181 nm) and nitrogen (174 nm) were monitored during the atomic plasma emission detection.

The GC-MS instrument used for the compound identification was the Agilent 6890N gas chromatograph connected to the 5973i MSD mass spectrometer (Agilent Technologies).

The GC column was a narrow-bore capillary with 180 μm, i.d. x 20 m DB-5MS (0.18 μm film thickness, Agilent Technologies, Wilmington, DE). The inlet head pressure was 12.5 psi for the helium flow of 0.7 ml/min. The system operated in the constant-flow mode. The temperature program was 50 °C for 2 min, followed by a ramp of 4 °C /min to 200 °C (hold time 1 min). Positive electron ionization (EI, 70 eV) mode was used with the scanning rate of 4.51 scans/sec over the mass range of 35-350 amu. The MSD transfer line temperature was set at 280 °C. The ion source and quadrupole temperatures were set at 230°C and 150°C, respectively. The TDSA-CIS sample introduction setting was identical to that described above in connection with the GC-AED system, except that the CIS trapping temperature was set at -80 °C.

Quantitative Evaluations and Statistical Analyses

As the basis for quantitative comparisons of urinary chromatographic profiles of the fresh urine collected on plates, the peak area integration was performed and peak areas were normalized by dividing with the peak area of internal standard (7-tridecanone) for each separated component. Either GC-MS total-ion chromatograms (TIC) or selected-ion chromatograms (SIC) obtained after the post-run modification of TICs were used for calculations. Normalized peak areas were statistically evaluated for the intact and castrated mouse groups. Student t test was employed for pairwise comparisons. The sulfur compound profiles from the GC-AED were compared in a qualitative manner between the intact and castrated males. The quantitative data obtained in this study for the male M. spicilegus urine samples was compared semi-quantitatively with the female M. spicilegus urine and qualitatively with the male M. domesticus urine data obtained in this laboratory previously.

RESULTS

Male M. spicilegus urinary volatile compound profiles by GC-MS featured more than one hundred components. Approximately 60 compounds showed sufficient spectral intensity and purity for the quantitative comparisons. Among these compounds, 46 were identified or partially identified while 10 remain unknown. The compound list is shown in Table 1.

The characteristic feature for the profiles from M. spicilegus was the prominent presence of and lactones, ketones, alcohols and acids. Characteristic urinary components found in M. domesticus are three dihydrofuran compounds (MW 126) originating from a puberty accelerating pheromone, 6-hydroxy-6-methyl-3-heptanone [START_REF] Novotny | A unique urinary constituent, 6-hydroxy-6-methyl-3-heptanone, has the puberty accelerating pheromone activity in mice[END_REF] and its lactol form (see Table 1 for 5,5-dimethyl-2-ethyl-4,5-dihydrofuran, Z-5,5-dimethyl-2ethylidenetetrahydrofuran, E-5,5-dimethyl-2-ethylidenetetrahydrofuran). These three furan derivatives were present in male M. spicilegus at the level comparable to the M. domesticus data reported earlier [START_REF] Harvey | Pattern of volatile compounds in dominant and subordinate male mouse urine[END_REF][START_REF] Novotny | Chemical identification of MHC-influenced volatile compounds in mouse urine. I: Quantitative proportions of major chemosignals[END_REF]. In the castrated M. spicilegus, the levels were significantly lower (P < 0.002) than in intact M. spicilegus males. Another M. domesticus pheromone compound, dehydro-exo-brevicomin, also present in female urine in small amounts [START_REF] Harvey | Pattern of volatile compounds in dominant and subordinate male mouse urine[END_REF][START_REF] Jemiolo | The t complex of the mouse: chemical characterization by urinary volatile profiles[END_REF], was found at lower levels in the intact and castrated male and also in female M. spicilegus urine (data not shown). Castration did not change dehydro-exo-brevicomin levels in M. spicilegus significantly in contrast to the suppressed levels of dehydro-exo-brevicomin in the samples from castrated M. domesticus [START_REF] Novotny | Chemical studies of the primer mouse pheromones[END_REF][START_REF] Harvey | Pattern of volatile compounds in dominant and subordinate male mouse urine[END_REF]. Trace levels of the dominant male mouse pheromone for M. domesticus, -farnesene [START_REF] Harvey | Pattern of volatile compounds in dominant and subordinate male mouse urine[END_REF], were detected in the intact male M. spicilegus urine, but were not seen in female or castrated animals. N-(methylthio)methylaniline was the only identified sulfur compound in the GC-MS TIC profile shown in M. spicilegus profiles, being more abundant in the intact male mouse urine when compared to the mouse urine of castrates (P<0.01). The representative structures of the main compounds distinguishing the two species are shown in Fig. 1. Castration of M. spicilegus males affected the volatile profiles in three ways: certain compounds, apparently under endocrine control, disappeared, while the levels of other compounds increased, and a set of previously undetected compounds became apparent (see Table 1). Fig. 2 illustrates the representative urinary volatile GC-MS TIC profiles for intact and castrated M. spicilegus males and intact male M. domesticus. In addition to -farnesene , castration removed 2-heptanone, -hexalactone, N-phenylformanilide and -octalactone from the set of urinary volatiles. Some compound levels decreased significantly after castration, including -hexalactone (P<0.02) and N-( methylthio)methylaniline ( P<0.01) as shown in Table 1. In contrast, after castration, levels of 2-coumaranone, a unique compound for M. spicilegus, increased significantly (P< 0.001) as demonstrated in Fig. 3. The absolute amounts of 2-coumaranone in castrated male urine were about 300 ng/ml  200 ng/ml (SD, n=6), while in the intact mouse urine, the levels were just 2 ng/ml  2 ng/ml (SD, n=14). In addition, other constituents, including several lactones, were found at higher levels, and new compounds such as coumarin and several late-eluting lactones appeared in the urine of castrated males (see Table 1). Fig. 4 shows a comparison of selected compound levels affected by castration. Individual variation for some of the compounds appeared relatively large (>70%, RSD), while some compounds varied only within the range of 12-30% (RSD) among the sampled individuals. Typically, the variation due to the sampling method was only 5-10% (RSD).

Qualitative comparisons in the GC-AED sulfur-selective profiles did not reveal any clear differences between intact and castrated animals. Relatively low-level (sub-picogram), sulfur-containing compounds (data not shown) were traced, but could not be identified structurally due to their extremely small representation.

Several of the previously reported M. domesticus urinary compounds (Novotny et al., 1990a;1990b[START_REF] Novotny | Chemical identification of MHC-influenced volatile compounds in mouse urine. I: Quantitative proportions of major chemosignals[END_REF] were also present in M. spicilegus profiles including three characteristic dihydrofurans. It is notable that the male dominance signaling pheromone compound 2-sec-butyl-4,5-dihydrothiazole and its "structural relative," 2-isopropyl-4,5dihydrothiazole for M. domesticus [START_REF] Novotny | Synthetic pheromones that promote inter-male aggression in mice[END_REF], were not detected in the M. spicilegus urine. Linear sulfur compounds such as dimethyl disulfide, bis(methylthio)methane and methyl(methylthio)methyl disulfide, typical for M. domesticus [START_REF] Novotny | Chemical identification of MHC-influenced volatile compounds in mouse urine. I: Quantitative proportions of major chemosignals[END_REF], were also absent in M. spicilegus GC-MS urinary volatile profiles. Furthermore, very few M. spicilegus urinary ketones were present compared to those identified in M. domesticus urine [START_REF] Novotny | Chemical identification of MHC-influenced volatile compounds in mouse urine. I: Quantitative proportions of major chemosignals[END_REF].

DISCUSSION

Since castrated male M. spicilegus did not show -farnesene 2-heptanone, -hexalactone, Nphenylformanilide and -octalactone among the urinary volatiles, it is suggested that the metabolic pathways involving these compounds may be under endocrine control. Behavioral tests using these compounds would be necessary to show what are the possible chemosignaling and physiological functions facilitated by these compounds for male and female M. spicilegus. Castration affected also most significantly the lactone structural types, thus demonstrating an endocrine feedback for the lactone biosynthesis. There is a strong possibility that some of the lactones may be mediators for chemical communication with the reproduction function. Furthermore, lactone levels were particularly varied among the individual intact males, so that the composition within the lactone compound profile could be related to individual recognition. In their pure form, many of the lactones exhibit fruit-or berry-like aromas [START_REF] Gatfield | Some aspects of the microbial production of flavor-active lactones with particular reference to decalactone[END_REF], which are relatively subtle odors in human perception, as opposed to the pungent smell in the M. domesticus male urine caused by the sulfur-containing compounds. The same unique lactone compounds were found in the female M. spicilegus urine in our qualitative screening (unpublished experiments).

Lactone biosynthesis involves C-18 hydroxyl fatty acids as precursors, which undergo -oxidation steps followed by lactonization [START_REF] Albrecht | Biosynthesis and biotechnological production of aliphatic and -lactones[END_REF]. Additionally, 9,10-oleic acid has been reported as a precursor for -dodecalactones in the yeast cultures [START_REF] Haffner | Biosynthesis of (R)--decanolactone in the yeast Sporobolomyces odorus[END_REF]. This may imply that the metabolism of fatty acids leading to urinary lactone end-products could play a prominent role in the M. spicilegus metabolism as opposed to M. domesticus which show very little presence of urinary lactones. The occurrence of urinary lactones has been previously reported in female and male pine voles (Microtus pinetorum) [START_REF] Boyer | Urinary volatile profiles of pine vole, Microtus pinetorum, and their endocrine dependency[END_REF]. In female pine voles, -octalactone exhibited the greatest urinary level changes among the volatile compounds after estrogen implantation or ovariectomy.

Other biological sources for lactones have been reported for insects and microorganisms, such as the cephalic gland -octalactone of the giant honeybee workers (Apis laboriosa) [START_REF] Blum | Chemical characters of two related species of giant honeybees (Apis dorsata and A. laboriosa): possible ecological significance[END_REF] and and -lactones emitted by marine Alphaproteobacteria [START_REF] Dickschat | The chafer pheromone buibuilactone and ant pyrazines are also produced by marine bacteria[END_REF]. In these two studies, lactones were hypothesized as potential chemical signaling compounds within the giant honeybee colony and bacterial culture, respectively.

Urinary ketone displays in intact male M. spicilegus were interestingly sparse (Table 1). M. domesticus urinary ketones (e.g., 2-heptanone, 6-methyl-5-hepten-3-one, 5-hepten-2one) have been previously found to correlate with the major histocompatibility complex (MHC) mouse haplotypes [START_REF] Novotny | Chemical identification of MHC-influenced volatile compounds in mouse urine. I: Quantitative proportions of major chemosignals[END_REF].

M. spicilegus originate from Eastern Europe. Several diagnostic genetic loci separate M. spicilegus and M. domesticus species [START_REF] Bonhomme | Biochemical diversity and evolution in the genus Mus[END_REF]reviewed in Sokolov et al., 1998). The genes in these loci code for several proteins, which are enzymatically active. The alcohol and malate dehydrogenases, esterases, carbonic anhydrase, mannose and glucose phosphate isomerases are among those proteins. Consequently, some of the distinguishing coded enzymes may impact the metabolic pathways leading to the excretion of urinary substances.

In addition to the lactones, urinary coumarin appears as an interesting "metabolic marker compound" for the M. spicilegus species. The presence of coumarin and abundance of 2-coumaranone in the urine of castrated M. spicilegus male also represent distinguishing metabolic pathways from M. domesticus. Coumarin could originate from different plants and herbs (reviewed in [START_REF] Bourgaud | Biosynthesis of coumarins in plants: a major pathway still to be unraveled for cytochrome P450 enzymes[END_REF]. In the mammalian systems, coumarin generally exhibits toxic effects and is oxidatively metabolized (detoxified) by the cytochrome P450 mono-oxygenase enzyme system in the liver microsomes [START_REF] Creaven | A spectrofluorimetric study of the 7-hydroxylation of coumarin by liver microsomes[END_REF][START_REF] Lewis | Species differences in coumarin metabolism: a molecular modeling evaluation of CYP2A interactions[END_REF]. The specific enzyme for coumarin elimination through 7-hydroxylation is CYP2A5 in mouse [START_REF] Miles | Identification of the human liver cytochrome P-450 responsible for coumarin 7-hydroxylase activity[END_REF]. The CYP2A5 enzyme is also known to be inhibited by lactones and 2-coumaranone [START_REF] Juvonen | Structural function of residue-209 in coumarin 7-hydroxylase (P450coh)[END_REF]2000). In a study within the M. domesticus strains, P450 enzymes have been found genetically altered among these strains [START_REF] Wood | Genetic regulation of coumarin hydroxylase activity in mice. Biochemical characterization of the enzyme from two inbred strains and their F 1 hybrid[END_REF]. Furthermore, a single autosomal gene locus Gpi-1 (glucose phosphate isomerase-1) was found responsible for the differential hydroxylase activity of P450 [START_REF] Wood | Genetic regulation of coumarin hydroxylase activity in mice. Evidence for single locus control on chromosome 7[END_REF]. Since Gpi-1 was also found as one of the distinguishing genetic loci between M. spicilegus and M. domesticus [START_REF] Bonhomme | Biochemical diversity and evolution in the genus Mus[END_REF], the vastly different urinary volatile profiles, as seen here, could be due in part to a differential P450 oxidase activity among other metabolic routes. For example, the distinguishing Es-2 loci controlling esterases have been found especially active in the kidney [START_REF] Ruddle | Esterase genetics in Mus musculus: expression, linkage, and polymorphism of locus Es-2[END_REF], and thus likely to affect some of the urinary metabolite excretion. However, separate genetic studies and metabolic mapping are necessary to link the genetic sources for the observed metabolic profile differences between M. spicilegus and M. domesticus, as exemplified by the urinary coumarin levels observed in this study. Furthermore, it seems desirable to investigate how could these genetically induced changes in the urinary volatile constituents facilitate chemical communication and social behavior in the "scent world" of the M. spicilegus species.

CONCLUSIONS

Quantitative comparisons of the urinary volatile profiles for male M. spicilegus mice reveal several compounds which have previously shown biological activity as male pheromones in the M. domesticus species. These similarities suggest that the two mouse species carry a certain genetic linkage that may be utilized in chemosignaling. On the other hand, the total absence of the prominent M. domesticus male aggression pheromone, 2-sec-butyl-4,5dihydrothiazole, in the M. spicilegus urine and the presence of unique and -lactones and coumarin seem to indicate that the species have developed distinctly separate metabolic pathways involving urinary constituents. As castration of M. spicilegus males removed hexalactone and -octalactone among the identified urinary constituents, their testosterone control suggests a possible involvement in chemical communication within the species.

Behavioral tests are in progress to define possible roles of several urinary volatile organic compounds in M. spicilegus chemical communication. dihydrofuran compounds, as indicated in Table 1.

  The experiments complied with the current French laws (authorization 93-0033 for C. Féron; Laboratory approval was secured from the Prefecture of Seine Saint Denis (prefectorial decree 02-2651), complying with the Association for the Study of Animal Behaviour/Animal Behaviour Society Guidelines for the Use of Animals in Research.

Fig. 1 .

 1 Fig.1. Distinguishing chemical structures of volatile compounds in M. spicilegus and M. domesticus male urine.

Fig 2 .

 2 Fig 2. Representative urinary (GC-MS total ion current) profiles of A: intact and B: castrated male M. spicilegus. C: A comparative profile for male intact M. domesticus.

Fig. 3 .

 3 Fig. 3. Urinary 2-coumaranone levels before and after castration.

Fig. 4 .

 4 Fig. 4. Effect of castration: compound levels significantly reduced in urine after castration. DHF (1-3)

Fig

  Fig.1.

  Fig.3

TABLE 1 .

 1 COMPARISON OF MALE MUS SPICILEGUS URINARY COMPOUNDS IN INTACT (I) AND CASTRATED ( C ) SAMPLESfor the compound level differences between intact (I) and castrated (C) male M. spicilegus urine (significance P < 0.05 accepted) ** unique urinay compounds for M.spicilegus, not found in M.domesticus

		Retention	Comparison	Student t-test
		time	of levels	P*
	Compound	(min)	I vs C*
	5,5-dimethyl-2-ethyl-4,5-		I > C	0.001
	dihydrofuran (DHF 1)	3.52
	Z-5,5-dimethyl-2-		I > C	0.001
	ethylidenetetrahydrofuran(DHF 2)	4.97
	2-heptanone	5.24	C=0
	E-5,5-dimethyl-2-		I > C	0.002
	ethylidenetetrahydrofuran (DHF 3)	5.72
	p-cymene	9.13	I > C	0.028
	3-octen-2-one	9.60
	dehydro-exo-brevicomin (DHB)	9.93
	-hexalactone**	9.99	I > C	0.023
	acetophenone	10.46	I > C	0.013
	o-toluidine	10.61
	-hexalactone**	11.37	C=0
	undecane	11.76	I > C	0.01
	nonanal	11.91	I > C	0.018
	phenylacetone	12.55
	a lactone**	13.38	I > C	0.001
	undecalactone**	13.63	I > C	0.032
	4-ethylphenol**	13.95	I > C	0.003
	a methyl toluate**	14.28
	octanoic acid	14.39
	decanal	15.42
	unidentified m/z 121**	15.63
	N-phenylformanilide	15.65	C=0
	unidentified m/z 140**	15.83
	2-coumaranone**	16.24	C > I	0.001
	a lactone**	16.96	I=0
	-octalactone**	17.16	C=0
	a lactone**	17.29	C > I	0.012
	nonanoic acid	17.64
	1-decanol	17.68
	-nonyl--valerolactone**	17.82
	a ketone	17.9	I > C	0.036
	indole	18.16
	coumarin**	18.35	I=0
	-nonenolactone**	19.77	C > I	0.048
	a lactone**	20.45	I=0
	an acid**	20.90
	undecanol	21.05
	N-( methylthio)methylaniline	21.92	I > C	0.01

Acknowledgements-This work was sponsored jointly by the Lilly Chemistry Alumni Chair and the Indiana METACyt Initiative of Indiana University, funded in part through a major grant from the Lilly Endowment, Inc. as well as the French National Center for Scientific Research and the French Ministry for Higher Education and Research (University of Paris 13).

We wish to thank Mr. Kevin Bruce in his assistance with the analyses and data collection, and Professor A. Hefetz for his comments on a previous version of the manuscript.


Jérôme Nevoux

Marie Teixeira

Say Viengchareun

Claudine Cosson

Daniel Butlen

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Evelyne Ferrary

Vasopressin, ATP and catecholamines differentially control potassium

  Figure1