Total Food Sustainability of the Agri-Food Chain
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Total Food Sustainability of the Agri-Food Chain
Edited by K. W. Waldron, G. K. Moates Institute of Food Research, Norwich, UK C. B. Faulds Centro de Investigaciones Biológicas (CSIC), Madrid, Spain
Preface The “Total Food” series of biennial, international conferences was initiated in 2004 by the Royal Society of Chemistry Food Group and the Institute of Food Research, Norwich. The aim of Total Food is to debate global research and development relevant to exploiting the whole food crop rather than the limited proportion that is consumed at present. For example, many vegetables are subjected to a high degree of trimming during post-harvest processing, and many parts including the outer leaves and stems are often disposed of by landfill. Much of the ‘wasted’ material is potentially food grade and may also contain important ingredients for further exploitation. The volumes involved are considerable. In the EC, over 500,000 tonnes of onion waste are disposed of per year. In some cases, up to 65% of food-grade material may be discarded during processing, and the EC-funded “AWARENET” report (2004) indicated that up to 222 million tonnes of food chain waste from all food processing sectors are produced annually across the EU. Since the Total Food series began, the issue of food security has become prominent. The increasing global population in conjunction with the use of crops for biofuel production mean that the more efficient exploitation of biomass will be required. The Total Food conferences are well placed to provide regular forums to highlight recent developments and to facilitate knowledge transfer between representatives of the agri-food (and increasingly non-food) industries, scientific research community, legal experts on foodrelated legislation and waste management, and consumer organisations. We gratefully acknowledge financial sponsorship from the following organisations without whose help the 2009 conference would not have been possible: the International AgriTechnology Centre, the Institute of Food Research, the Food & Health Network, Norwich Research Park and Achor International. Information and downloads: Total Food 2009 Conference details, abstracts and presentations may be obtained from the website: http://www.ifr.ac.uk/totalfood2009/ Total Food 2004: Presentations, abstracts and the complete Proceedings Volume may be downloaded from the website http://www.totalfood2004.com/
K.W. Waldron G.K. Moates C.B. Faulds
Contents
Key Drivers A socio-economic perspective on co-product exploitation B. Gremmen, P. van Haperen and J. Lamerichs
3
Value added products - Plants Food applications of novel ingredients from agro-based sustainable sources J.L. Bialek, D. Jarvis and P. Lopez-Sanchez
17
Improving the textural characteristics of brewer’s spent grain breads by combination of sour dough and different enzymes V. Stojceska and P. Ainsworth
27
Effect of packaging conditions on shelf-life of bologna sausages made with orange juice wastewater and oregano essential oil M. Viuda-Martos, Y. Ruiz-Navajas, J. Fernández-López, E. Sendra, E. Sayas-Barbera and J.A. Pérez-Álvarez
32
Effects of industrial processing on content and properties of dietary fibre of strawberry wastes P. Torres, F.J. López-Andréu, G. Torres, M. Vidriales, R.M. Esteban, E. Mollá and M.A. Martín Cabrejas
38
Formulation and acceptability studies of high fibre cookies made from pink guava (Psidium guajava) decanter / agro waste H. Chek Zaini, H. Zaiton, C.W. Zanariah and N. Sakinah
44
Extraction of antioxidant compounds from apple pomace H.H. Wijngaard and N. Brunton
53
Extracting novel foam and emulsion stability enhancers from brewers’ grain F.A. Husband, A. Jay, C.B. Faulds, K.W. Waldron and P.J. Wilde
58
Biological production of vanillin from ferulic acid obtained from wheat bran hydrolyzates D. Di Gioia, L. Sciubba, M. Ruzzi and F. Fava
64
Methanolic extract of Cistus ladaniferus as a source of phenolic antioxidants for use in foods M. Amensour, M. Viuda-Martos, E. Sendra, J. Abrini, J.A. Pérez-Álvarez and J. Fernández-López
70
viii
Contents
Contents
ix
x
Contents
Asparagus fibres as reinforcing materials for developing 100% biodegradable packaging S. Jaramillo-Carmona, R. Guillén, C. Escrig-Rondan, J.M. Fuentes-Alventosa, G. Rodríguez, A. Lama, A. Jiménez-Araujo, J. Fernández-Bolaños and R. Rodríguez-Arcos
224
Increasing protein extraction yield from duckweed (Lemna obscura) with an ammonia treatment L. Urribarrí, J. Ríos and A. Ferrer
229
Diffusion of bioactive peptides from chitosan-based edible films – effects of temperature and peptides molecular weight A.C. Pinheiro, A.I. Bourbon, M.A.C. Quintas, C. Rocha, J.A. Teixeira and A.A. Vicente
233
Functional properties of Gleditsia triacanthos seeds extracts and their incorporation into galactomannan films for food applications M.A. Cerqueira, B.W.S. Souza, J.T. Martins, J.A. Teixeira and A.A. Vicente
238
An investigation on the effect of formulation and extrusion temperature on physico-chemical characteristics of tomato-enriched snacks Z. Dehghan-Shoar, A. Hardacre, G. Meerdink and C.S. Brennan
244
Subject Index
249
4
Total Food
Key Drivers
5
6
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Key Drivers
7
8
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included questions that addressed the feasibility and significance of REPRO’s contribution to reprocess food waste and whether the trajectories REPRO is working on (BSG, trimmings) are best suited to reach the project’s goals. Similar to what was said previously, for some parts REPRO can be considered a project which aim is to exemplify processes and tools, but for another part the actual significance or feasibility cannot be determined yet. Critical remarks are made with regard to whether the trajectories are best suited to reach the project’s goals, and to feasibility in terms of economical feasibility (the costs involved). 3.2 Delphi study among Stakeholders In the Delphi study among stakeholders we used almost the method as in the Delphi study among Repro members. The differences were that we used ranking questions as well, the “no opinion” option was left out, and the questionnaire was restricted to 16 questions. From the 54 invited candidates only 16 eventually responded. The main result was that the diverging viewpoints were concerned with the relative importance of communication towards consumers and product-origin-information, the relative importance of increased quality and hazard control and, the appropriateness of existing reprocessing technology. Food safety and quality control was seen as the core issue in relation to consumer trust and acceptance, but communication was considered important by others as a precaution to win trust and handle negative reactions. The idea for consumers to know that food was produced from waste was seen as rather threatening, while reprocessing itself was not seen as risky, just uneconomical for the moment. This in contrast to remarks that the origin of the by product itself plays an important role. Reprocessing inside the production chain is seen as more acceptable than outside, reprocessing vegetable matter more consumer acceptable than animal products. Legislation and quality control was not perceived to cause greater waste streams, although it alternately was regarded as an obstacle to further utilize by-products. Waste handling legislation was also indicated to form a possible incentive in stimulating reprocessing techniques. Where some reported a lack of technology for reprocessing, others indicated economic incentives were missing to employ existing known reprocessing technology. Others again were pointing out that the existing techniques were lacking an outlook for profitable and acceptable products. Overall in the study, it appeared that although there was sufficient interest in discussing quality aspects of reprocessing of food products, the stakeholders in majority choose techniques for volume reduction and water recycling –not resulting in food products- as the most promising areas for reprocessing.
4 FOCUS GROUP DISCUSSIONS Focus groups are used to discover what people think and feel with the intention to define the concept of satisfaction, identify the relevant ingredients of satisfaction and discover the conditions or circumstances that influence satisfaction. Characteristics of a focus group: people who possess certain characteristics and provide qualitative data in a focused discussion to help understand the topic of interest. Four focus groups were conducted of respectively 8, 10, 10 and 9 respondents, a total of 37 respondents. After completing the fourth focus group theoretical saturation was reached: the range of ideas was heard, answers overlapped. Moreover, it was not doable to conduct more focus groups in the Netherlands as well as any abroad with the time and resources available. The duration of
Key Drivers
9
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Key Drivers
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Key Drivers
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18
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Value Added Products - Plants
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plateau. A 2hr-time scale was used for the further study of the particle size effect on the gel formation. 30000 <1mm <2mm
25000
G' 2h (Pa)
20000 15000 10000 5000
N TT 10
% 80
20
%
TT +
TT + % 40
0%
N TT
N TT % 60
% 50
%
TT +
40 TT + % 60
50 %
20 TT +
10 80 %
N TT
N TT
N TT %
0%
TT
0
Figure 2 Phase volume and particle size effect (insoluble solids) on the resultant gel strength (G’24). Conditions: Time sweep (1 Pa shear stress, 1Hz frequency), temperature 30°C. Figure 2 shows both the effect of changing the NTT/TT ratio and the effect of reducing the particle size. The results are presented as a function of elastic modulus after 2hrs of incubation at 30°C. Error bars indicate a relatively large variability of the measurements. This could be due to both environmental (change in temperature, humidity, etc.) and equipment related problems, but it is clear that the nature of the raw material is the main reason. Studies of living material, such as vegetables, are usually subject to this variability problem. As a consequence, our analysis focuses more on trends than absolute results. The same general trends were observed for both particle sizes considered in this study. NTT onion purée had a greater storage modulus than its TT counterpart and the largest value of G’2h was obtained for 60% (w/w) TT + 40% (w/w) NTT onion mixtures. As shown in Figure 2, a small amount of NTT onion added to TT onion purée was sufficient to obtain a significant increase in G’2h, whereas in the opposite scenario this trend was less pronounced. From these observations, it seems that NTT/ TT onion purées act on the resultant gel strength differently. The reduction of particle size results in a decrease of the G’2h values for all ratios studied. However this decrease was less significant for 100% TT onion purée (about 2000 Pa) than for 100% NTT onion purée (about 5000 Pa).
22
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2.1 Preparation of BSG The source of BSG was based on brewing of barley hops and is the remains after wort mashing and straining, before fermentation of the beer. The BSG was refrigerated immediately after collection and then dried the following day at 150°C for 4 hours in a Teknotronic reel oven (Teknigas Ltd, Sussex, England) to a moisture content of 5-7%. The dried sample was milled using 0.5 mesh screens, packed and then sealed in polyethylene bags and kept at room temperature until use. The composition of dried BSG was 20.30% protein, 53.39% fibre, 8.32% fat and 10.76% carbohydrates. 2.2 Preparation of sour dough breads The dough was prepared with mixing sour dough, soaker dough, wheat flour, water, salt and yeast. Sour dough was prepared with mixing BÖCKER TK Starter, wheat flour and water. Soaker dough was made by mixing BSG and water. Mixing of the ingredients was carried out in a Hobart NCM mixer (Process Plant and Machinery Ltd., UK) at 1st speed. The resulting dough was divided into three 300g pieces, moulded by hand and placed into pre-greased 454g tins. The dough was proofed 55 min at 30°C and 85% humidity and baked in a Tektronic reel oven (Teknigas, Ltd., Sussex, England) for 20 min at 230°C. The following bread types were studied: • Three control sourdough breads (SBB) with the substitution levels of BSG at 0, 10,
20 and 30%. • Three sourdough breads using Lipopan Extra enzyme (SLE) with the substitution
levels of BSG at 0, 10, 20 and 30%. • Three sourdough breads using Pentopan enzyme (SPE) with the substitution levels of
BSG at 0, 10, 20 and 30%. • Three sourdough breads using a mixture of Pentopan + Celluclast enzymes (SPCE)
with the substitution levels of BSG at 0, 10, 20 and 30%. • Three sourdough breads using a mixture of Lipopan Extra and Maxlife 85 enzymes
(SLME) with the substitution levels of BSG at 0, 10, 20 and 30%. • Three sourdough breads using a mixture of Pentopan and Maxlife 85 (SPME)
enzymes with the substitution levels of BSG at 0, 10, 20 and 30%. 2.3 Bread evaluation Quality analysis of fresh bread samples was carried out by measuring weight, volume (determined by seed displacement in a loaf volume meter) and specific loaf volume. Crumb firmness was measured using a texture analyzer (TA-XT2i, Stable Micro Systems, Surrey, UK) at 1, 5 and 8 days to assess potential shelf-life of the breads. Bread crumb firmness during storage was determined as maximum compression force4. The loaves were sliced by hand into equal slice thickness of 25 mm thick. The crust slices of the loaves were discarded. Nine bread slices originating from three loaves were measured and results were expressed as mean values.
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2.5 Polyphenol analysis 2.5.1 FCR assay. Total phenolic content of ethanolic apple pomace extracts was assessed using a modified version of the Folin-Ciocalteu assay.13 An aqueous gallic acid solution (200 mg / l) was used to create a standard curve. For the analysis, 100 µl of sample extract or gallic acid standard, 100 µl of methanol, 100 µl of Folin-Ciocalteu reagent and 700 µl of Na2CO3 were aliquoted into a 1.5 ml micro-centrifuge tube. The samples were vortexed immediately and the tubes were incubated in the dark for 20 min at room temperature. After incubation all samples were centrifuged at 13,000 rpm for 3 min. The absorbance of the supernatant was then measured at 735 nm using a spectrophotometer (UV-1700 Pharma Spec, Shimadzu, Japan). The results were expressed in mg gallic acid equivalent / 100 g dry weight (mg GAE / 100 g DW). 2.5.2 HPLC-DAD. HPLC-analysis was performed on a Varian Pro Star (Varian Inc., Walnut Creek, USA) chromatograph system, equipped with a module 335 diode array detector with an absorbance detection range between 190 to 950 nm. Separations were conducted on a Zorbax SB C18, 5µm, 150 x 4.6mm column (Agilent Technologies, Dublin, Ireland). The gradient profile was based on a method of Tsao and Yang.14 Acetic acid in 2 mM sodium acetate (final pH 2.55, v/v) was used as eluent A and 100% acetonitrile was used as eluent B; a gradient profile was used. Hydroxybenzoic acids, dihydrochalcones, procyanidins and flavanols were monitored at a wavelength of 280 nm, hydroxycinnamic acid derivatives at 320 nm, and flavonols at 360 nm. For quantification and identification purposes standard curves of analytes of interest were prepared using authenticated standards. Levels of chlorogenic acid were expressed in µmol chlorogenic acid / 100 g DW apple pomace, total flavonol level was expressed in µmol rutin / 100 g DW apple pomace and the level of phloretin glycosides was expressed in µmol phlorizin / 100 g DW apple pomace. 2.6 Experimental Design Response surface methodology (RSM) was used to optimise antioxidant activity and polyphenol extraction from apple pomace. A Central Composite design was used for both conventional extraction and PLE. The independent variables were coded at five levels (-α, -1, 0, 1, α) and was using five central points. Design Expert 7.1.3 (Stat-Ease Inc., Minneapolis, USA) was used for designing experiments and statistical data analysis (ANOVA). Experimental data were analysed to fit a second order polynomial model. Models and regression coefficients were considered significant when p-values were lower than 0.05.
3 RESULTS AND DISCUSSION 3.1 Conventional extraction The main polyphenols present in solid-liquid extracts from apple pomace are chlorogenic acid, flavonols, and phloretin glucosides.10 Catechins and procyanidins and other cinnamic acids such as caffeic acid are present in very small amounts. Anthocyanins are absent from the extracts. Similar results have been reported by other authors, when extracting apple pomace with ethanol.15 Analysis of these polyphenols, DPPH values and total phenols as response values by RSM has revealed that second order polynomial models for these
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Value Added Products - Fish, Meat and Dairy
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132
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Value Added Products - Fish, Meat and Dairy
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Value Added Products - Fish, Meat and Dairy
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136
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Value Added Products - Fish, Meat and Dairy
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Value Added Products - Fish, Meat and Dairy
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140
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Value Added Products - Fish, Meat and Dairy
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Value Added Products - Fish, Meat and Dairy
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144
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Value Added Products - Fish, Meat and Dairy
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146
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Measuring Sustainability
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Measuring Sustainability
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Integrated Approaches - Process and Chain
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Integrated Approaches - Process and Chain
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Integrated Approaches - Process and Chain
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Integrated Approaches - Process and Chain
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Integrated Approaches - Process and Chain
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Energy Recovery and Technologies for Water Recovery and Recycling
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Energy Recovery and Technologies for Water Recovery and Recycling
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Energy Recovery and Technologies for Water Recovery and Recycling
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Energy Recovery and Technologies for Water Recovery and Recycling
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180
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Energy Recovery and Technologies for Water Recovery and Recycling
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Energy Recovery and Technologies for Water Recovery and Recycling
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Energy Recovery and Technologies for Water Recovery and Recycling
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Energy Recovery and Technologies for Water Recovery and Recycling
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190
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Energy Recovery and Technologies for Water Recovery and Recycling
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192
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Energy Recovery and Technologies for Water Recovery and Recycling
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Energy Recovery and Technologies for Water Recovery and Recycling
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Energy Recovery and Technologies for Water Recovery and Recycling
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Table 1 Physical and chemical parameters of Rum DSW before (unfermented) and after 9 days fermentation with A. niger. DSW1: DSW from Reunion Island.
Data from Literature
Measured parameters
Unfermented DSW1*
DSW1 after fermentation*
References
pH
4.2 - 5.9
4.6
5.4
7
Total solids (g.L-1)
4 - 106
189
201
8, 9
10.7 - 28.9
41.4
43.2
10
1.1 - 10.9
12.9
81.9
8, 11
32 - 90
146.5
53
11, 12
7.9
11.8
11.3
13
Ashes (g.L-1) -1
Suspended Solids (SS) (g.L ) COD (g.L-1) Carbon/Nitrogen ratio (C/N)
* Data are average of 3 independent experiments.
The more important differences were found for Total Solids and COD. DSW1 showed in average values for both parameters about two times higher than published data. These differences are probably the results from differences in sugar cane strains, soils composition and distillation processes. The high content of suspended solids present in unfermented DSW sample (12.92 g.L-1) is due to the yeast present during alcohol fermentation. After 9 days fermentation, very high amount of Suspended Solids were detected (82 g.L-1 in average). This value includes filterable biomass produced during the fermentation process. While biomass increased, COD decreased proportionally by means of 64 %. Preliminary results also indicated that Absorbance of DSW measured at 270 nm also strongly decreased during fermentation (from 310 to 145 units).
3 CONCLUSIONS Analyses of the physical and chemical characteristics of DSW from Reunion Island confirmed that this effluent have a high polluting load. Parameters like COD, SS, and absorbance confirmed this conclusion. Our results clearly show that Aspergillus niger can perfectly grow on unsupplemented DSW and we obtained on average about 70 g.L-1 of biomass in the better case. Incubation of A. niger in DSW media offers biotreatment potentialities since COD was decreased by a factor 3 during fermentation. According to biomass production, we can also conclude that the fungus has a strong potential for bioproduction. This process still has to be optimized but at this stage of the work, it offers very interesting perspectives for industrial applications. Both aspects, namely bioproduction and bioremediation will be further study with other strains alone or combined in mixed cultures. References 1 2 3 4
K. Kida, S. Morimura, N. Abe and Y. Sonoda, Proc. Biochem. 1995, 30, 125. A.C. Wilkie, K.J. Riedesel and J.M. Owens, Biomass and Bioenergy 2000, 19, 63. S. Karhaman and O. Yesilada, Folia Microbiol., 2003, 48, 525. F.J. Fitzgibbon, P. Nigam, D. Singh and R. Marchant, J. Basic Microbiol., 1995, 35, 293.
Energy Recovery and Technologies for Water Recovery and Recycling
199
Energy Recovery and Technologies for Water Recovery and Recycling
201
202
Total Food
Energy Recovery and Technologies for Water Recovery and Recycling
203
Energy Recovery and Technologies for Water Recovery and Recycling
205
206
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Energy Recovery and Technologies for Water Recovery and Recycling
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212
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Bulk Products for Food, Feed and Non-food Uses
213
214
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Bulk Products for Food, Feed and Non-food Uses
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216
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Bulk Products for Food, Feed and Non-food Uses
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218
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220
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Bulk Products for Food, Feed and Non-food Uses
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222
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Bulk Products for Food, Feed and Non-food Uses
223
Bulk Products for Food, Feed and Non-food Uses
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226
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Bulk Products for Food, Feed and Non-food Uses
227
228
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230
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Bulk Products for Food, Feed and Non-food Uses
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232
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234
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Bulk Products for Food, Feed and Non-food Uses
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0,7
1
0,5
0,8
0,4
Mt/Meq
Mt/Meq
Flores et al.
1,2
0,6
0,3 Experimental
0,2
0,6 Experimental 0,4
Predicted
Predicted
0,2
0,1
Predicted Global
Predicted Global 0 0
50
100
150 200 Time (min)
250
300
350
0 0
2000
4000
6000 8000 Time (min)
10000
12000
Bulk Products for Food, Feed and Non-food Uses
237
Bulk Products for Food, Feed and Non-food Uses
239
240
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Bulk Products for Food, Feed and Non-food Uses
241
242
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Bulk Products for Food, Feed and Non-food Uses
243
Bulk Products for Food, Feed and Non-food Uses
245
246
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Subject Index
accelerated solvent extraction, see extraction, accelerated solvent almond by-products, 86 ammonia treatment, 229 animal feed, 219 anthocyanin, see dyes, anthocyanins, 211 anti-inflammatory compounds, 141 antimicrobial effect, 86, 136 antioxidants, 53, 90, 119, 136, 140, 240 apple, peel, 90 pomace, 53 arabinoxylans, 58 asparagus, see fibre, asparagus Aspergillus niger, 196 aquaculture waste, 200 bacterial phytase, see phytase beer, 61 bioactive peptides, see peptides, bioactive biodegradable packaging, 224 biofuel, 181 biofouling, see fouling, biobiohydrogen, 171, 204 biomethane, 171, 200 blue whiting, see fish waste, blue whiting blumenol, 72 Bologna sausages, see sausages, Bologna botifarró, see sausages, botifarró bovine muscle, 134 bovine offal, 134 bran, rice, 97 wheat, 64, 206 bread making, 27 breakfast cereals, see extrusion, breakfast cereals brewers spent grain, 1, 27, 58, 150 broccoli, 114 cabbage juice, 11, 153 carrot, 114
catechins, 55, 89 cellulase activity, 181 inhibitory effort of alcohols, 184 cell wall sugars, 39, 118 cheese manufacture, 130 chitosan films, 233 chlorogenic acid, 55, 94 Cistus ladaniferus, 70 cleaning cycle optimization, 132 colonic fermentation, see fermentation, colonic cookies, 44 co-product exploitation, 3, 149 corn cob, 206 cream cheese, 74 crosslinking, oxidative, 109 dairy processing, 129 Delphi studies, 7 dietary fibre, see fibre, dietary diffusion modelling, 234 digestion, anaerobic, 171, 200 in vitro, 86 dihydrocalchones, 94 duckweed, 229 dyes, anthocyanins, 211 flavonoid, 211 natural, 211 tannins, 211 edible films, 233, 238 emulsion stability enhancers, 58 Enterobacter sakazakii, 96 energy efficiency, 158 environmental impact, 151 enzymes, in bread making, 27 degradation of meat proteins, 134 epicatechin, 89
250
essential oil, Cistus ladaniferus, 70 oregano, 32 extraction, accelerated solvent, 53 ethanolic, 120 methanolic, 70, 120 using organic acids, 142 pressurised liquid, 53 protein, 229 extrusion, biodegradable packaging, 225 breakfast cereals, 105 snack products, 10, 150 tomato-enriched snacks, 244 feed production, 219 fermentation, colonic, 87 photo-, 204 simultaneous saccharification and fermentation, 181 thermophilic, 204 ferulic acid, 64, 111 fibre, asparagus, 224 dietary, 11, 27, 38, 47, 80 duckweed, 230 lemon, 74 orange, 74, 80 fish waste, blue whiting, 142 gilthead, 201 mackerel, 138, 142 sea bass, 201 trout, 201 flavan-3-ols, 94 flavonoids, 71, 90 flavonoid dyes, see dyes, flavonoid flavonols, 55 flavonol glycosides, 90 foam stability enhancers, 58 fouling, bio-, 132 protein/mineral deposits, 132 FTIR spectroscopy, 117 galactomannan films, 238 gelatines, 142 emulsifying properties, 144
Subject Index
foaming properties, 145 gel strength, 145 gelation, 17 gel strength, 25, 145 gilthead, see fish waste, gilthead Gleditsia triacanthos, 238 Gracilaria birdiae, 119 Gracilaria cornea, 119 grapes, pressed, 211 greenhouse gas emissions, 151 guava waste, 44 hydrogen, see biohydrogen in vitro digestion, see digestion, in vitro isorhamnetin, 89 laccase, 109 Last Minute Market model, 162 LCA, see life cycle assessment Lemna obscura, 229 lemon fibre see fibre, lemon life cycle assessment, 149, 158 lignocellulosic biomass, 181 limonene, 72 lycopene, 244 mackerel, see fish waste, mackerel manure, 175 marine waste, 138, 142 mathematical optimisation, 188 meat waste, 134 methane, see biomethane microbiological analysis, 77, 114 molasses, 196, 206 naringenin, 89 onion peel, 211 puree, 17 orange fibre see fibre, orange oregano see essential oil, oregano oxidative crosslinking, see crosslinking, oxidative pectin, 11, 17 pectin methylesterase, 18 peptides, bioactive, from meat, 134
Subject Index
251
