83a5c0c08ee84f9193dd63442008b9aa  FNS_a2014v5p1746.pdf
021335c2de140fba96a07458b2d81e7123e76d94  FNS_a2014v5p1746.pdf
69efc6db85913acf4f2dbe4c00480f0d9bb202759048591aae77be6d53d9cae8  FNS_a2014v5p1746.pdf

Title:           Enhacement of the Viability of Lactobacillus plantarum during the Preservation and Storage Process Based on the Response Surface Methodology  
Subject:         Objective: The Response Surface Methodology (RSM) is a commonly used system to optimize cell viability of probiotic strains when they are subjected to different preservation and storage processes. Methods and Results: To determine the optimal levels of incorporation of several cry oprotectants (skim milk, sucrose and trehalose) in the freeze-drying process of Lactobacillus plantarum, a range of experiments based on a Rotational Central Composite design (CCD) were conducted. The results were adjusted to a quadratic model, resulting in the presence of interaction between the different variables. Solving a regression equation, we obtained the optimum concentrations of cryoprotective agents: 24.06% milk powder, 6.22% sucrose, 5.63% trehalose. To visualize the interactions between the three variables involved in the study, Design Expert® software was used. Conclusions: The analysis reveals that while trehalose has a direct effect on the viability of L. plantarum, skim milk and sucrose exert quadratic effects. There are also interactions between cryoprotectants, which emphasize the synergies produced between milk and sucrose and between sucrose and trehalose, which allows maintaining the viability of L. plantarum. Significance and Impact of the Study: The addition of new oligosaccharides as trehalose in premixtures for functional feed can maintain the viability of L. plantarum during longer periods of time, ensuring the proper administration of probiotics to their destinations.
Keywords:        Cryoprotectants, Freeze-Drying Process, Probiotics, Response Surface Methodology, Central Composite Design
Author:          Girme Gisela, Arosemena Esteban Leonardo, Perez Lucia, Vera Rodrigo, Grau Eduard, Calvo M. Angeles
Creator:         Acrobat PDFMaker 11 Word 版
Producer:        Adobe PDF Library 11.0
CreationDate:    Fri Sep 19 08:50:46 2014 CEST
ModDate:         Fri Sep 19 09:02:51 2014 CEST
Custom Metadata: yes
Metadata Stream: yes
Tagged:          no
UserProperties:  no
Suspects:        no
Form:            none
JavaScript:      no
Pages:           11
Encrypted:       no
Page size:       595.32 x 807.96 pts
Page rot:        0
File size:       4847968 bytes
Optimized:       no
PDF version:     1.6


name                                 type              encoding         emb sub uni object ID
------------------------------------ ----------------- ---------------- --- --- --- ---------
GDEMGL+Calibri-Bold                  TrueType          WinAnsi          yes yes yes    307  0
WOEEWF+Calibri                       TrueType          WinAnsi          yes yes yes    309  0
TimesNewRomanPSMT                    TrueType          WinAnsi          no  no  no     311  0
MUZRMZ+Calibri-Italic                TrueType          WinAnsi          yes yes yes    313  0
Tahoma                               TrueType          WinAnsi          no  no  no     315  0
CFZJCT+Cambria-Bold                  TrueType          WinAnsi          yes yes yes    317  0
CFZJCT+Cambria-BoldItalic            TrueType          WinAnsi          yes yes yes    319  0
TimesNewRomanPS-BoldMT               TrueType          WinAnsi          no  no  no     321  0
SQZBSN+Cambria                       TrueType          WinAnsi          yes yes yes    204  0
SQZBSN+Cambria-Italic                TrueType          WinAnsi          yes yes yes    202  0
IWZTIH+SymbolMT                      CID TrueType      Identity-H       yes yes yes    215  0
TimesNewRomanPSMT                    CID TrueType      Identity-H       yes no  yes    210  0
TimesNewRomanPS-ItalicMT             CID TrueType      Identity-H       yes no  yes    209  0
TimesNewRomanPS-BoldMT               CID TrueType      Identity-H       yes no  yes    208  0
TimesNewRomanPS-ItalicMT             TrueType          WinAnsi          no  no  no     206  0
TimesNewRomanPS-BoldItalicMT         TrueType          WinAnsi          no  no  no     220  0
ArialMT                              TrueType          WinAnsi          no  no  no     221  0


Jhove (Rel. 1.28.0, 2023-05-18)
 Date: 2025-10-15 03:53:47 CEST
 RepresentationInformation: FNS_a2014v5p1746.pdf
  ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16)
  LastModified: 2025-10-14 14:21:08 CEST
  Size: 4847968
  Format: PDF
  Version: 1.6
  Status: Well-Formed and valid
  SignatureMatches:
   PDF-hul
  MIMEtype: application/pdf
  PDFMetadata: 
   Objects: 0
   FreeObjects: 18
   IncrementalUpdates: 0
   DocumentCatalog: 
    PageLayout: OneColumn
    PageMode: UseOutlines
    Outlines: 
     Item: 
      Title: Enhacement of the Viability of Lactobacillus plantarum during the Preservation and Storage Process Based on the Response Surface Methodology  
     Item: 
      Title: Abstract
     Item: 
      Title: Keywords
     Item: 
      Title: 1. Introduction
     Item: 
      Title: 2. Materials and Methods
      Children: 
       Item: 
        Title: 2.1. Bacterial Culture and Bacterial Culture Medium
       Item: 
        Title: 2.2. Cryoprotective Medium and Preparation of Suspension
       Item: 
        Title: 2.3. Sample Preparation
       Item: 
        Title: 2.4. Lyophilization
       Item: 
        Title: 2.5. Determining the Viability of Microorganisms
        Children: 
         Item: 
          Title: 2.5.1. Determination of Viability Pre-Lyophilization
         Item: 
          Title: 2.5.2. Determining the Conservation of Viability over Time
     Item: 
      Title: 3. Results 
      Children: 
       Item: 
        Title: 3.1. Central Composite Rotational Design Analysis Using Response Surface Methodology
       Item: 
        Title: 3.2. Effects of Protective Agents
     Item: 
      Title: 4. Discussion
     Item: 
      Title: References
   Filters: 
    FilterPipeline: FlateDecode
    FilterPipeline: DCTDecode
   Images: 
    Image: 
     NisoImageMetadata: 
      FormatName: image/png
      ImageWidth: 2755
      ImageHeight: 13
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Filter: FlateDecode
     Decode: 0, 255
     Name: X
    Image: 
     NisoImageMetadata: 
      FormatName: image/png
      ImageWidth: 88
      ImageHeight: 31
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Filter: FlateDecode
     Decode: 0, 255
     Name: X
    Image: 
     NisoImageMetadata: 
      FormatName: image/png
      ImageWidth: 272
      ImageHeight: 108
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Filter: FlateDecode
     Name: X
    Image: 
     NisoImageMetadata: 
      FormatName: image/png
      ImageWidth: 2755
      ImageHeight: 13
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Filter: FlateDecode
     Decode: 0, 255
     Name: X
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 241
      ImageHeight: 60
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: X
    Image: 
     NisoImageMetadata: 
      FormatName: image/png
      ImageWidth: 1300
      ImageHeight: 912
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Filter: FlateDecode
     Name: X
    Image: 
     NisoImageMetadata: 
      FormatName: image/png
      ImageWidth: 1300
      ImageHeight: 902
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Filter: FlateDecode
     Name: X
    Image: 
     NisoImageMetadata: 
      FormatName: image/png
      ImageWidth: 1300
      ImageHeight: 877
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Filter: FlateDecode
     Name: X
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im20
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im31
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im21
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im30
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im24
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im14
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im25
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im22
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im23
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im17
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im28
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im18
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im29
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im26
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im27
    Image: 
     NisoImageMetadata: 
      FormatName: image/jpg
      CompressionScheme: JPEG
      ImageWidth: 396
      ImageHeight: 538
      BitsPerSample: 8
      BitsPerSampleUnit: integer
     Name: Im19
   Fonts: 
    Type0: 
     Font: 
      BaseFont: TimesNewRomanPS-BoldMT
      Encoding: Identity-H
      ToUnicode: true
     Font: 
      BaseFont: TimesNewRomanPS-ItalicMT
      Encoding: Identity-H
      ToUnicode: true
     Font: 
      BaseFont: TimesNewRomanPSMT
      Encoding: Identity-H
      ToUnicode: true
     Font: 
      BaseFont: IWZTIH+SymbolMT
      Encoding: Identity-H
      ToUnicode: true
    TrueType: 
     Font: 
      BaseFont: TimesNewRomanPS-BoldMT
      FirstChar: 0
      LastChar: 255
      FontDescriptor: 
       FontName: TimesNewRomanPS-BoldMT
       Flags: Serif, Nonsymbolic
       FontBBox: -558, -307, 2000, 1026
      Encoding: WinAnsiEncoding
     Font: 
      BaseFont: SQZBSN+Cambria-Italic
      FontSubset: true
      FirstChar: 32
      LastChar: 116
      FontDescriptor: 
       FontName: SQZBSN+Cambria-Italic
       Flags: Serif, Nonsymbolic, Italic
       FontBBox: -1105, -279, 1222, 1047
       FontFile2: true
      Encoding: WinAnsiEncoding
      ToUnicode: true
     Font: 
      BaseFont: SQZBSN+Cambria
      FontSubset: true
      FirstChar: 32
      LastChar: 115
      FontDescriptor: 
       FontName: SQZBSN+Cambria
       Flags: Serif, Nonsymbolic
       FontBBox: -1475, -2463, 2868, 3117
       FontFile2: true
      Encoding: WinAnsiEncoding
      ToUnicode: true
     Font: 
      BaseFont: TimesNewRomanPS-ItalicMT
      FirstChar: 0
      LastChar: 255
      FontDescriptor: 
       FontName: TimesNewRomanPS-ItalicMT
       Flags: Serif, Nonsymbolic, Italic
       FontBBox: -498, -307, 1333, 1023
      Encoding: WinAnsiEncoding
     Font: 
      BaseFont: GDEMGL+Calibri-Bold
      FontSubset: true
      FirstChar: 32
      LastChar: 119
      FontDescriptor: 
       FontName: GDEMGL+Calibri-Bold
       Flags: Nonsymbolic
       FontBBox: -519, -306, 1240, 971
       FontFile2: true
      Encoding: WinAnsiEncoding
      ToUnicode: true
     Font: 
      BaseFont: WOEEWF+Calibri
      FontSubset: true
      FirstChar: 32
      LastChar: 243
      FontDescriptor: 
       FontName: WOEEWF+Calibri
       Flags: Nonsymbolic
       FontBBox: -503, -307, 1240, 964
       FontFile2: true
      Encoding: WinAnsiEncoding
      ToUnicode: true
     Font: 
      BaseFont: TimesNewRomanPSMT
      FirstChar: 0
      LastChar: 255
      FontDescriptor: 
       FontName: TimesNewRomanPSMT
       Flags: Serif, Nonsymbolic
       FontBBox: -568, -307, 2000, 1007
      Encoding: WinAnsiEncoding
     Font: 
      BaseFont: MUZRMZ+Calibri-Italic
      FontSubset: true
      FirstChar: 32
      LastChar: 117
      FontDescriptor: 
       FontName: MUZRMZ+Calibri-Italic
       Flags: Nonsymbolic, Italic
       FontBBox: -725, -276, 1260, 1014
       FontFile2: true
      Encoding: WinAnsiEncoding
      ToUnicode: true
     Font: 
      BaseFont: Tahoma
      FirstChar: 0
      LastChar: 255
      FontDescriptor: 
       FontName: Tahoma
       Flags: Nonsymbolic
       FontBBox: -600, -216, 1516, 1034
      Encoding: WinAnsiEncoding
     Font: 
      BaseFont: TimesNewRomanPS-BoldItalicMT
      FirstChar: 0
      LastChar: 255
      FontDescriptor: 
       FontName: TimesNewRomanPS-BoldItalicMT
       Flags: Serif, Nonsymbolic, Italic
       FontBBox: -547, -307, 1401, 1032
      Encoding: WinAnsiEncoding
     Font: 
      BaseFont: CFZJCT+Cambria-Bold
      FontSubset: true
      FirstChar: 32
      LastChar: 174
      FontDescriptor: 
       FontName: CFZJCT+Cambria-Bold
       Flags: Serif, Nonsymbolic
       FontBBox: -1110, -299, 1373, 1047
       FontFile2: true
      Encoding: WinAnsiEncoding
      ToUnicode: true
     Font: 
      BaseFont: ArialMT
      FirstChar: 0
      LastChar: 255
      FontDescriptor: 
       FontName: ArialMT
       Flags: Nonsymbolic
       FontBBox: -665, -325, 2000, 1006
      Encoding: WinAnsiEncoding
     Font: 
      BaseFont: CFZJCT+Cambria-BoldItalic
      FontSubset: true
      FirstChar: 32
      LastChar: 117
      FontDescriptor: 
       FontName: CFZJCT+Cambria-BoldItalic
       Flags: Serif, Nonsymbolic, Italic
       FontBBox: -1110, -299, 1378, 1047
       FontFile2: true
      Encoding: WinAnsiEncoding
      ToUnicode: true
    CIDFontType2: 
     Font: 
      BaseFont: TimesNewRomanPS-ItalicMT
      CIDSystemInfo: 
       Registry: Adobe
       Registry: Identity
       Supplement: 0
      FontDescriptor: 
       FontName: TimesNewRomanPS-ItalicMT
       Flags: Serif, Symbolic, Italic
       FontBBox: -498, -307, 1333, 1023
       FontFile2: true
     Font: 
      BaseFont: TimesNewRomanPSMT
      CIDSystemInfo: 
       Registry: Adobe
       Registry: Identity
       Supplement: 0
      FontDescriptor: 
       FontName: TimesNewRomanPSMT
       Flags: Serif, Symbolic
       FontBBox: -568, -307, 2000, 1007
       FontFile2: true
     Font: 
      BaseFont: TimesNewRomanPS-BoldMT
      CIDSystemInfo: 
       Registry: Adobe
       Registry: Identity
       Supplement: 0
      FontDescriptor: 
       FontName: TimesNewRomanPS-BoldMT
       Flags: Serif, Symbolic
       FontBBox: -558, -307, 2000, 1026
       FontFile2: true
     Font: 
      BaseFont: IWZTIH+SymbolMT
      CIDSystemInfo: 
       Registry: Adobe
       Registry: Identity
       Supplement: 0
      FontDescriptor: 
       FontName: IWZTIH+SymbolMT
       Flags: Symbolic
       FontBBox: 0, -220, 1113, 1005
       FontFile2: true
   XMP: <x:xmpmeta xmlns:x="adobe:ns:meta/" x:xmptk="Adobe XMP Core 5.4-c005 78.147326, 2012/08/23-13:03:03        ">
   <rdf:RDF xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#">
      <rdf:Description rdf:about=""
            xmlns:xmp="http://ns.adobe.com/xap/1.0/"
            xmlns:xmpMM="http://ns.adobe.com/xap/1.0/mm/"
            xmlns:dc="http://purl.org/dc/elements/1.1/"
            xmlns:pdf="http://ns.adobe.com/pdf/1.3/"
            xmlns:pdfx="http://ns.adobe.com/pdfx/1.3/"
            xmlns:photoshop="http://ns.adobe.com/photoshop/1.0/">
         <xmp:ModifyDate>2014-09-19T15:02:51+08:00</xmp:ModifyDate>
         <xmp:CreateDate>2014-09-19T14:50:46+08:00</xmp:CreateDate>
         <xmp:MetadataDate>2014-09-19T15:02:51+08:00</xmp:MetadataDate>
         <xmp:CreatorTool>Acrobat PDFMaker 11 Word 版</xmp:CreatorTool>
         <xmpMM:DocumentID>uuid:591344fd-9046-4cbc-905d-e410487bbe29</xmpMM:DocumentID>
         <xmpMM:InstanceID>uuid:bf6d07f7-1467-49c8-bd86-36d7d5479661</xmpMM:InstanceID>
         <xmpMM:subject>
            <rdf:Seq>
               <rdf:li>294</rdf:li>
            </rdf:Seq>
         </xmpMM:subject>
         <dc:format>application/pdf</dc:format>
         <dc:title>
            <rdf:Alt>
               <rdf:li xml:lang="x-default">Enhacement of the Viability of Lactobacillus plantarum during the Preservation and Storage Process Based on the Response Surface Methodology  </rdf:li>
            </rdf:Alt>
         </dc:title>
         <dc:description>
            <rdf:Alt>
               <rdf:li xml:lang="x-default">Objective: The Response Surface Methodology (RSM) is a commonly used system to optimize cell viability of probiotic strains when they are subjected to different preservation and storage processes. Methods and Results: To determine the optimal levels of incorporation of several cry oprotectants (skim milk, sucrose and trehalose) in the freeze-drying process of Lactobacillus plantarum, a range of experiments based on a Rotational Central Composite design (CCD) were conducted. The results were adjusted to a quadratic model, resulting in the presence of interaction between the different variables. Solving a regression equation, we obtained the optimum concentrations of cryoprotective agents: 24.06% milk powder, 6.22% sucrose, 5.63% trehalose. To visualize the interactions between the three variables involved in the study, Design Expert® software was used. Conclusions: The analysis reveals that while trehalose has a direct effect on the viability of L. plantarum, skim milk and sucrose exert quadratic effects. There are also interactions between cryoprotectants, which emphasize the synergies produced between milk and sucrose and between sucrose and trehalose, which allows maintaining the viability of L. plantarum. Significance and Impact of the Study: The addition of new oligosaccharides as trehalose in premixtures for functional feed can maintain the viability of L. plantarum during longer periods of time, ensuring the proper administration of probiotics to their destinations.</rdf:li>
            </rdf:Alt>
         </dc:description>
         <dc:creator>
            <rdf:Seq>
               <rdf:li>Girme Gisela, Arosemena Esteban Leonardo, Perez Lucia, Vera Rodrigo, Grau Eduard, Calvo M. Angeles</rdf:li>
            </rdf:Seq>
         </dc:creator>
         <pdf:Producer>Adobe PDF Library 11.0</pdf:Producer>
         <pdf:Keywords>Cryoprotectants, Freeze-Drying Process, Probiotics, Response Surface Methodology, Central Composite Design</pdf:Keywords>
         <pdfx:SourceModified>D:20140919064935</pdfx:SourceModified>
         <pdfx:Company/>
         <photoshop:headline>
            <rdf:Seq>
               <rdf:li>Objective: The Response Surface Methodology (RSM) is a commonly used system to optimize cell viability of probiotic strains when they are subjected to different preservation and storage processes. Methods and Results: To determine the optimal levels of incorporation of several cry oprotectants (skim milk, sucrose and trehalose) in the freeze-drying process of Lactobacillus plantarum, a range of experiments based on a Rotational Central Composite design (CCD) were conducted. The results were adjusted to a quadratic model, resulting in the presence of interaction between the different variables. Solving a regression equation, we obtained the optimum concentrations of cryoprotective agents: 24.06% milk powder, 6.22% sucrose, 5.63% trehalose. To visualize the interactions between the three variables involved in the study, Design Expert® software was used. Conclusions: The analysis reveals that while trehalose has a direct effect on the viability of L. plantarum, skim milk and sucrose exert quadratic effects. There are also interactions between cryoprotectants, which emphasize the synergies produced between milk and sucrose and between sucrose and trehalose, which allows maintaining the viability of L. plantarum. Significance and Impact of the Study: The addition of new oligosaccharides as trehalose in premixtures for functional feed can maintain the viability of L. plantarum during longer periods of time, ensuring the proper administration of probiotics to their destinations.</rdf:li>
            </rdf:Seq>
         </photoshop:headline>
      </rdf:Description>
   </rdf:RDF>
</x:xmpmeta>
   Pages: 
    Page: 
     Sequence: 1
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 216, 729, 343, 740
      Annotation: 
       Subtype: Link
       Rect: 54, 694, 218, 729
      Annotation: 
       Subtype: Link
       Rect: 283, 42, 447, 53
      Annotation: 
       Subtype: Link
       Rect: 471, 724, 537, 750
      Annotation: 
       Subtype: Link
       Rect: 111, 481, 216, 493
      Annotation: 
       Subtype: Link
       Rect: 84, 411, 274, 422
    Page: 
     Sequence: 2
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 141, 668, 157, 680
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 156, 668, 172, 680
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 311, 668, 328, 680
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 326, 668, 343, 680
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 428, 619, 444, 632
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 372, 583, 388, 595
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 354, 546, 375, 559
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 408, 486, 429, 498
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 519, 449, 540, 461
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 84, 437, 105, 449
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 289, 425, 311, 437
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 309, 425, 330, 437
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 278, 401, 300, 413
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 298, 401, 319, 413
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 286, 328, 307, 340
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 113, 291, 134, 303
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 460, 267, 481, 279
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 84, 243, 105, 255
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 103, 243, 124, 255
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 511, 206, 532, 218
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 346, 194, 367, 206
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 476, 182, 497, 194
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 517, 170, 538, 182
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 517, 146, 538, 158
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 181, 121, 202, 133
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 248, 97, 270, 109
       ActionDest: 1
    Page: 
     Sequence: 3
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 303, 710, 324, 722
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 281, 428, 302, 440
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 301, 428, 323, 440
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 479, 416, 500, 428
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 498, 416, 519, 428
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 483, 404, 520, 416
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 242, 285, 278, 297
       ActionDest: 1
    Page: 
     Sequence: 4
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 385, 650, 422, 662
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 460, 560, 481, 572
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 177, 476, 213, 488
       ActionDest: 1
    Page: 
     Sequence: 5
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 87, 284, 124, 295
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 169, 236, 205, 248
       ActionDest: 1
    Page: 
     Sequence: 6
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 105, 434, 141, 446
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 247, 275, 284, 287
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 326, 89, 366, 101
       ActionDest: 1
    Page: 
     Sequence: 7
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 328, 224, 369, 236
       ActionDest: 1
      Annotation: 
       Subtype: Link
       Rect: 222, 188, 262, 200
       ActionDest: 1
    Page: 
     Sequence: 8
    Page: 
     Sequence: 9
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 105, 698, 291, 712
      Annotation: 
       Subtype: Link
       Rect: 191, 674, 369, 688
      Annotation: 
       Subtype: Link
       Rect: 105, 639, 283, 653
      Annotation: 
       Subtype: Link
       Rect: 282, 615, 450, 629
      Annotation: 
       Subtype: Link
       Rect: 214, 580, 365, 594
      Annotation: 
       Subtype: Link
       Rect: 105, 546, 279, 560
      Annotation: 
       Subtype: Link
       Rect: 219, 511, 378, 525
      Annotation: 
       Subtype: Link
       Rect: 332, 487, 493, 501
      Annotation: 
       Subtype: Link
       Rect: 105, 452, 298, 466
      Annotation: 
       Subtype: Link
       Rect: 105, 428, 493, 442
      Annotation: 
       Subtype: Link
       Rect: 123, 393, 309, 407
      Annotation: 
       Subtype: Link
       Rect: 123, 334, 316, 348
      Annotation: 
       Subtype: Link
       Rect: 105, 299, 278, 313
      Annotation: 
       Subtype: Link
       Rect: 105, 240, 291, 254
      Annotation: 
       Subtype: Link
       Rect: 105, 216, 393, 230
      Annotation: 
       Subtype: Link
       Rect: 105, 182, 286, 195
      Annotation: 
       Subtype: Link
       Rect: 105, 147, 284, 161
      Annotation: 
       Subtype: Link
       Rect: 193, 123, 377, 137
      Annotation: 
       Subtype: Link
       Rect: 370, 98, 503, 112
    Page: 
     Sequence: 10
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 105, 708, 291, 722
      Annotation: 
       Subtype: Link
       Rect: 341, 674, 509, 688
      Annotation: 
       Subtype: Link
       Rect: 299, 639, 467, 653
      Annotation: 
       Subtype: Link
       Rect: 253, 605, 445, 618
    Page: 
     Sequence: 11
     Annotations: 
      Annotation: 
       Subtype: Link
       Rect: 28, 765, 122, 804
      Annotation: 
       Subtype: Link
       Rect: 470, 784, 561, 800
      Annotation: 
       Subtype: Link
       Rect: 151, 625, 291, 640
      Annotation: 
       Subtype: Link
       Rect: 75, 479, 169, 609
      Annotation: 
       Subtype: Link
       Rect: 424, 480, 518, 608
      Annotation: 
       Subtype: Link
       Rect: 307, 480, 402, 608
      Annotation: 
       Subtype: Link
       Rect: 192, 480, 285, 608
      Annotation: 
       Subtype: Link
       Rect: 74, 329, 169, 458
      Annotation: 
       Subtype: Link
       Rect: 191, 329, 285, 458
      Annotation: 
       Subtype: Link
       Rect: 307, 329, 402, 458
      Annotation: 
       Subtype: Link
       Rect: 424, 330, 517, 458
      Annotation: 
       Subtype: Link
       Rect: 75, 179, 169, 308
      Annotation: 
       Subtype: Link
       Rect: 192, 179, 286, 308
      Annotation: 
       Subtype: Link
       Rect: 307, 179, 402, 308
      Annotation: 
       Subtype: Link
       Rect: 424, 179, 518, 308
      Annotation: 
       Subtype: Link
       Rect: 75, 29, 169, 158
      Annotation: 
       Subtype: Link
       Rect: 191, 29, 286, 157
      Annotation: 
       Subtype: Link
       Rect: 307, 29, 402, 158
      Annotation: 
       Subtype: Link
       Rect: 423, 29, 518, 158
      Annotation: 
       Subtype: Link
       Rect: 36, 622, 136, 641
  Checksum: 53d07823
   Type: CRC32
  Checksum: 83a5c0c08ee84f9193dd63442008b9aa
   Type: MD5
  Checksum: 021335c2de140fba96a07458b2d81e7123e76d94
   Type: SHA-1
  Checksum: 69efc6db85913acf4f2dbe4c00480f0d9bb202759048591aae77be6d53d9cae8
   Type: SHA-256