d5fcbdd380a782432c307835bf5a372e nucacires_a2025m11v53n20pgkaf1134.pdf 73658e8bf1e1580b722c555d967f8a648eea2dde nucacires_a2025m11v53n20pgkaf1134.pdf dfe5882a44b3c63ba6b9e15b4e78a47ef75e295d752f0fd00d5eab19aafd47df nucacires_a2025m11v53n20pgkaf1134.pdf Title: A versatile plug and play bacterial chassis based on a fast-growing Mycoplasma species Subject: DOI: 10.1093/nar/gkaf1134 Nucleic Acids Research, 53, 20, 00-00-2025. Abstract: An easy engineerable bacterial chassis opens a spectrum of biomedical applications such as antigen or drug delivery. However, most cell-walled bacteria are readily recognized and cleared by the immune system and encode various effector proteins potentially interacting with the target species. Mycoplasma feriruminatoris, a cell-wall-less minimal organism with excellent growth attributes, was genetically engineered to stably and almost seamlessly incorporate foreign DNA fragments into the chromosome in a single step using an orthogonal recombinase system. Using this mechanism, we characterized and developed promoters and proved their functionality, including a kill-switch based on a temperature-sensitive promoter that drives expression of a toxin from a toxin–antitoxin system. Additionally, we designed a streptavidin-based functional dynamic display system at the surface of the bacterium. The engineered strains have great potential such as to deliver vaccine antigens to different livestock species among other biomedical applications. Keywords: Author: Inäbnit Jana, Démoulins Thomas, Pramateftaki Paraskevi, Bourgeon Marilou, Labroussaa Fabien, Jores Jörg, Torres-Puig Sergi Creator: OUP Producer: Acrobat Distiller 25.0 (Windows); modified using iTextSharp.LGPLv2.Core 3.7.4.0 CreationDate: Sat Nov 8 06:51:38 2025 CET ModDate: Mon Jun 15 11:01:02 2026 CEST Custom Metadata: yes Metadata Stream: yes Tagged: yes UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 612 x 792 pts (letter) Page rot: 0 File size: 2575692 bytes Optimized: no PDF version: 1.4 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- BIOCKI+SabonLTStd-Roman Type 1C Custom yes yes yes 73 0 BIOFEB+UniversLTStd-LightObl Type 1C Custom yes yes yes 134 0 BIOGAN+SabonLTStd-Bold Type 1C Custom yes yes yes 497 0 BIOCKJ+dummy-space Type 1C WinAnsi yes yes no 78 0 BIOCKK+UniversLTStd-CnObl Type 1C Custom yes yes yes 2064 0 BIOCKL+UniversLTStd-Cn Type 1C Custom yes yes yes 2065 0 BIOCKM+UniversLTStd-Bold Type 1C Custom yes yes yes 79 0 BIOCKN+UniversLTStd-BoldCn Type 1C Custom yes yes yes 2066 0 BIODFK+UniversLTStd-BoldObl Type 1C Custom yes yes yes 2067 0 BIOFCA+MTSY Type 1C Custom yes yes yes 80 0 BIOFEA+UniversLTStd-Light Type 1C Custom yes yes yes 135 0 GXBEAA+ArialMT TrueType WinAnsi yes yes no 1 0 BIOGKL+SabonLTStd-Italic Type 1C Custom yes yes yes 74 0 BIOIPN+UniversLTStd Type 1C Custom yes yes yes 75 0 BIOJAO+EURM10 Type 1C Custom yes yes yes 76 0 BIOJAP+RMTMI Type 1C Custom yes yes yes 77 0 BIOJIN+UniversLTStd-Obl Type 1C Custom yes yes yes 101 0 BIPBNL+EURM7 Type 1C Custom yes yes yes 199 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-06-16 04:08:31 CEST RepresentationInformation: nucacires_a2025m11v53n20pgkaf1134.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-06-15 11:01:58 CEST Size: 2575692 Format: PDF Version: 1.4 Status: Well-Formed, but not valid SignatureMatches: PDF-hul ErrorMessage: Invalid indirect destination - referenced object 'FN1' cannot be found ID: PDF-HUL-149 ErrorMessage: Invalid indirect destination - referenced object 'COR1' cannot be found ID: PDF-HUL-149 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'FN1' cannot be found ID: PDF-HUL-122 Offset: 2207141 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'COR1' cannot be found ID: PDF-HUL-122 Offset: 2207141 MIMEtype: application/pdf Profile: Tagged PDF PDFMetadata: Objects: 2141 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: true NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: OneColumn PageMode: UseThumbs Language: English Outlines: Item: Title: Introduction Destination: section.1.1.0 Item: Title: Materials and methods Destination: section.2.2.0 Item: Title: Results Destination: section.3.20.0 Item: Title: Discussion Destination: section.4.29.0 Item: Title: Acknowledgements Destination: section.5.30.0 Item: Title: Supplementary data Destination: section.6.31.0 Item: Title: Conflict of interest Destination: section.7.32.0 Item: Title: Funding Destination: section.8.33.0 Item: Title: Data availability Destination: section.9.34.0 Item: Title: References Destination: section.10.34.0 Info: Title: A versatile plug and play bacterial chassis based on a fast-growing Mycoplasma species Author: Inäbnit Jana, Démoulins Thomas, Pramateftaki Paraskevi, Bourgeon Marilou, Labroussaa Fabien, Jores Jörg, Torres-Puig Sergi Subject: DOI: 10.1093/nar/gkaf1134 Nucleic Acids Research, 53, 20, 00-00-2025. Abstract: An easy engineerable bacterial chassis opens a spectrum of biomedical applications such as antigen or drug delivery. However, most cell-walled bacteria are readily recognized and cleared by the immune system and encode various effector proteins potentially interacting with the target species. Mycoplasma feriruminatoris, a cell-wall-less minimal organism with excellent growth attributes, was genetically engineered to stably and almost seamlessly incorporate foreign DNA fragments into the chromosome in a single step using an orthogonal recombinase system. Using this mechanism, we characterized and developed promoters and proved their functionality, including a kill-switch based on a temperature-sensitive promoter that drives expression of a toxin from a toxin–antitoxin system. Additionally, we designed a streptavidin-based functional dynamic display system at the surface of the bacterium. The engineered strains have great potential such as to deliver vaccine antigens to different livestock species among other biomedical applications. Keywords: Creator: OUP Producer: Acrobat Distiller 25.0 (Windows); modified using iTextSharp.LGPLv2.Core 3.7.4.0 CreationDate: Sat Nov 08 06:51:38 CET 2025 ModDate: Mon Jun 15 11:01:02 CEST 2026 ID: 0xe1ba0ba1de374e63630ba560adc74bed, 0xa829cb7d403b1b0b1cbf7281900ba47d Filters: FilterPipeline: FlateDecode FilterPipeline: DCTDecode Images: Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1327 ImageHeight: 602 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1402 ImageHeight: 643 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1399 ImageHeight: 844 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Image: NisoImageMetadata: FormatName: image/png ImageWidth: 974 ImageHeight: 1156 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Filter: FlateDecode Image: NisoImageMetadata: FormatName: image/png ImageWidth: 1067 ImageHeight: 1376 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Filter: FlateDecode Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1064 ImageHeight: 1114 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 967 ImageHeight: 1302 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Fonts: Type1: Font: BaseFont: BIOJIN+UniversLTStd-Obl FontSubset: true FirstChar: 45 LastChar: 121 FontDescriptor: FontName: BIOJIN+UniversLTStd-Obl Flags: Symbolic, Italic FontBBox: -174, -272, 1145, 947 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOFEB+UniversLTStd-LightObl FontSubset: true FirstChar: 45 LastChar: 121 FontDescriptor: FontName: BIOFEB+UniversLTStd-LightObl Flags: Symbolic, Italic FontBBox: -151, -274, 1144, 957 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOFEA+UniversLTStd-Light FontSubset: true FirstChar: 2 LastChar: 142 FontDescriptor: FontName: BIOFEA+UniversLTStd-Light Flags: Symbolic FontBBox: -151, -250, 1000, 957 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIPBNL+EURM7 FontSubset: true FirstChar: 2 LastChar: 181 FontDescriptor: FontName: BIPBNL+EURM7 Flags: Serif, Symbolic FontBBox: -5, -243, 1229, 720 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOCKI+SabonLTStd-Roman FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: BIOCKI+SabonLTStd-Roman Flags: Serif, Symbolic FontBBox: -165, -228, 1000, 944 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOGKL+SabonLTStd-Italic FontSubset: true FirstChar: 2 LastChar: 121 FontDescriptor: FontName: BIOGKL+SabonLTStd-Italic Flags: Symbolic, Italic FontBBox: -146, -228, 1059, 927 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOIPN+UniversLTStd FontSubset: true FirstChar: 2 LastChar: 133 FontDescriptor: FontName: BIOIPN+UniversLTStd Flags: Symbolic FontBBox: -168, -250, 992, 947 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOJAO+EURM10 FontSubset: true FirstChar: 2 LastChar: 181 FontDescriptor: FontName: BIOJAO+EURM10 Flags: Serif, Symbolic FontBBox: -32, -243, 1060, 720 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOJAP+RMTMI FontSubset: true FirstChar: 2 LastChar: 62 FontDescriptor: FontName: BIOJAP+RMTMI Flags: Symbolic, Italic FontBBox: 0, -213, 987, 680 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOCKJ+dummy-space FontSubset: true FirstChar: 32 LastChar: 32 FontDescriptor: FontName: BIOCKJ+dummy-space Flags: Nonsymbolic FontBBox: 0, 0, 0, 0 FontFile3: true Encoding: WinAnsiEncoding Font: BaseFont: BIOCKM+UniversLTStd-Bold FontSubset: true FirstChar: 2 LastChar: 246 FontDescriptor: FontName: BIOCKM+UniversLTStd-Bold Flags: Symbolic, ForceBold FontBBox: -167, -250, 995, 938 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOFCA+MTSY FontSubset: true FirstChar: 2 LastChar: 215 FontDescriptor: FontName: BIOFCA+MTSY Flags: Symbolic FontBBox: 0, -954, 1043, 796 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOCKK+UniversLTStd-CnObl FontSubset: true FirstChar: 65 LastChar: 117 FontDescriptor: FontName: BIOCKK+UniversLTStd-CnObl Flags: Symbolic, Italic FontBBox: -172, -270, 1064, 989 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOGAN+SabonLTStd-Bold FontSubset: true FirstChar: 58 LastChar: 118 FontDescriptor: FontName: BIOGAN+SabonLTStd-Bold Flags: Serif, Symbolic FontBBox: -163, -228, 1000, 968 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOCKL+UniversLTStd-Cn FontSubset: true FirstChar: 40 LastChar: 242 FontDescriptor: FontName: BIOCKL+UniversLTStd-Cn Flags: Symbolic FontBBox: -166, -250, 1000, 989 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIOCKN+UniversLTStd-BoldCn FontSubset: true FirstChar: 66 LastChar: 121 FontDescriptor: FontName: BIOCKN+UniversLTStd-BoldCn Flags: Symbolic, ForceBold FontBBox: -83, -250, 1000, 969 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BIODFK+UniversLTStd-BoldObl FontSubset: true FirstChar: 77 LastChar: 121 FontDescriptor: FontName: BIODFK+UniversLTStd-BoldObl Flags: Symbolic, Italic, ForceBold FontBBox: -173, -270, 1148, 938 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true TrueType: Font: BaseFont: GXBEAA+ArialMT FontSubset: true FirstChar: 32 LastChar: 121 FontDescriptor: FontName: GXBEAA+ArialMT Flags: Nonsymbolic FontBBox: -664, -324, 2000, 1039 FontFile2: true Encoding: WinAnsiEncoding XMP: 2025-11-08T05:51:38Z 2026-06-15T09:01:02+00:00 2026-06-15T09:01:02+00:00 OUP Acrobat Distiller 25.0 (Windows); modified using iTextSharp.LGPLv2.Core 3.7.4.0 VoR 10.1093/nar/gkaf1134 1362-4962 Nucleic Acids Research journal 53 10.1093/nar/gkaf1134 https://doi.org/10.1093/nar/gkaf1134 © The Author(s) 2025. Published by Oxford University Press.This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com. All Rights Reserved application/pdf doi:10.1093/nar/gkaf1134 © The Author(s) 2025. Published by Oxford University Press.This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com. A versatile plug and play bacterial chassis based on a fast-growing Mycoplasma species DOI: 10.1093/nar/gkaf1134 Nucleic Acids Research, 53, 20, 00-00-2025. Abstract: An easy engineerable bacterial chassis opens a spectrum of biomedical applications such as antigen or drug delivery. However, most cell-walled bacteria are readily recognized and cleared by the immune system and encode various effector proteins potentially interacting with the target species. Mycoplasma feriruminatoris, a cell-wall-less minimal organism with excellent growth attributes, was genetically engineered to stably and almost seamlessly incorporate foreign DNA fragments into the chromosome in a single step using an orthogonal recombinase system. Using this mechanism, we characterized and developed promoters and proved their functionality, including a kill-switch based on a temperature-sensitive promoter that drives expression of a toxin from a toxin–antitoxin system. Additionally, we designed a streptavidin-based functional dynamic display system at the surface of the bacterium. The engineered strains have great potential such as to deliver vaccine antigens to different livestock species among other biomedical applications. Inäbnit Jana Démoulins Thomas Pramateftaki Paraskevi Bourgeon Marilou Labroussaa Fabien Jores Jörg Torres-Puig Sergi Oxford University Press Inäbnit Jana, Démoulins Thomas, Pramateftaki Paraskevi, Bourgeon Marilou, Labroussaa Fabien, Jores Jörg, Torres-Puig Sergi uuid:c7fa63c1-6d70-3940-a70f-0aab7959c0e7 uuid:8fa2390d-3320-4945-ba82-7289c8bcd70f True Pages: Page: Label: 1 Annotations: Annotation: Subtype: Link Rect: 46, 740, 161, 750 Annotation: Subtype: Link Rect: 111, 635, 117, 645 Destination: 665 Annotation: Subtype: Link Rect: 223, 635, 230, 645 Destination: 665 Annotation: Subtype: Link Rect: 354, 630, 364, 644 Annotation: Subtype: Link Rect: 362, 635, 368, 645 Destination: 665 Annotation: Subtype: Link Rect: 469, 630, 478, 644 Annotation: Subtype: Link Rect: 476, 635, 483, 645 Destination: 665 Annotation: Subtype: Link Rect: 144, 620, 150, 630 Destination: 665 Annotation: Subtype: Link Rect: 150, 620, 156, 630 Destination: 665 Annotation: Subtype: Link Rect: 156, 620, 163, 630 Destination: 665 Annotation: Subtype: Link Rect: 222, 620, 228, 630 Destination: 665 Annotation: Subtype: Link Rect: 228, 620, 234, 630 Destination: 665 Annotation: Subtype: Link Rect: 235, 620, 241, 630 Destination: null Annotation: Subtype: Link Rect: 241, 620, 248, 630 Destination: null Annotation: Subtype: Link Rect: 342, 615, 352, 629 Annotation: Subtype: Link Rect: 350, 620, 356, 630 Destination: 665 Annotation: Subtype: Link Rect: 356, 620, 363, 630 Destination: 665 Annotation: Subtype: Link Rect: 363, 620, 369, 630 Destination: null Annotation: Subtype: Link Rect: 370, 620, 376, 630 Destination: null Annotation: Subtype: Link Rect: 224, 528, 294, 538 Annotation: Subtype: Link Rect: 264, 519, 333, 529 Annotation: Subtype: Link Rect: 49, 73, 204, 83 Annotation: Subtype: Link Rect: 272, 64, 334, 74 Annotation: Subtype: Link Rect: 46, 45, 149, 55 Thumb: true Page: Label: 2 Annotations: Annotation: Subtype: Link Rect: 172, 679, 179, 691 Destination: 305 Annotation: Subtype: Link Rect: 97, 668, 104, 680 Destination: 305 Annotation: Subtype: Link Rect: 101, 613, 108, 625 Destination: 305 Annotation: Subtype: Link Rect: 236, 547, 243, 559 Destination: 305 Annotation: Subtype: Link Rect: 216, 404, 224, 416 Destination: 305 Annotation: Subtype: Link Rect: 109, 382, 116, 394 Destination: 305 Annotation: Subtype: Link Rect: 273, 382, 280, 394 Destination: 305 Annotation: Subtype: Link Rect: 141, 305, 148, 317 Destination: 333 Annotation: Subtype: Link Rect: 248, 283, 255, 295 Destination: 333 Annotation: Subtype: Link Rect: 61, 239, 73, 251 Destination: 333 Annotation: Subtype: Link Rect: 288, 228, 300, 240 Destination: 333 Annotation: Subtype: Link Rect: 146, 173, 158, 185 Destination: 333 Annotation: Subtype: Link Rect: 173, 118, 185, 130 Destination: 333 Annotation: Subtype: Link Rect: 189, 118, 201, 130 Destination: 333 Annotation: Subtype: Link Rect: 97, 107, 109, 119 Destination: 333 Annotation: Subtype: Link Rect: 182, 96, 194, 108 Destination: 333 Annotation: Subtype: Link Rect: 82, 74, 94, 86 Destination: 333 Annotation: Subtype: Link Rect: 246, 74, 259, 86 Destination: 333 Annotation: Subtype: Link Rect: 99, 63, 111, 75 Destination: 333 Annotation: Subtype: Link Rect: 61, 52, 73, 64 Destination: 333 Annotation: Subtype: Link Rect: 409, 617, 421, 629 Destination: 333 Annotation: Subtype: Link Rect: 318, 561, 565, 586 Destination: 333 Annotation: Subtype: Link Rect: 391, 330, 490, 342 Annotation: Subtype: Link Rect: 331, 281, 432, 293 Annotation: Subtype: Link Rect: 394, 237, 401, 249 Destination: 333 Annotation: Subtype: Link Rect: 322, 160, 334, 172 Destination: 333 Annotation: Subtype: Link Rect: 337, 160, 349, 172 Destination: 333 Annotation: Subtype: Link Rect: 498, 77, 510, 89 Destination: 333 Thumb: true Page: Label: 3 Annotations: Annotation: Subtype: Link Rect: 97, 650, 197, 662 Annotation: Subtype: Link Rect: 46, 617, 128, 629 Annotation: Subtype: Link Rect: 280, 580, 287, 592 Destination: 333 Annotation: Subtype: Link Rect: 247, 356, 254, 368 Destination: 333 Annotation: Subtype: Link Rect: 63, 323, 149, 335 Annotation: Subtype: Link Rect: 50, 103, 62, 115 Destination: 333 Annotation: Subtype: Link Rect: 495, 584, 502, 596 Destination: 333 Annotation: Subtype: Link Rect: 335, 404, 347, 416 Destination: 333 Annotation: Subtype: Link Rect: 336, 99, 348, 111 Destination: 333 Annotation: Subtype: Link Rect: 519, 88, 531, 100 Destination: 333 Annotation: Subtype: Link Rect: 346, 44, 358, 56 Destination: 333 Annotation: Subtype: Link Rect: 428, 33, 528, 45 Thumb: true Page: Label: 4 Annotations: Annotation: Subtype: Link Rect: 239, 705, 251, 717 Destination: 333 Annotation: Subtype: Link Rect: 104, 683, 112, 695 Destination: 333 Annotation: Subtype: Link Rect: 114, 683, 127, 695 Destination: 333 Annotation: Subtype: Link Rect: 94, 263, 193, 275 Annotation: Subtype: Link Rect: 187, 214, 194, 226 Destination: 333 Annotation: Subtype: Link Rect: 184, 55, 191, 67 Destination: 333 Annotation: Subtype: Link Rect: 195, 55, 207, 67 Destination: 333 Annotation: Subtype: Link Rect: 451, 610, 463, 622 Destination: 333 Annotation: Subtype: Link Rect: 358, 360, 370, 372 Destination: 333 Annotation: Subtype: Link Rect: 512, 338, 524, 350 Destination: 333 Thumb: true Page: Label: 5 Annotations: Annotation: Subtype: Link Rect: 335, 445, 440, 455 Annotation: Subtype: Link Rect: 173, 110, 180, 122 Destination: 120 Annotation: Subtype: Link Rect: 146, 99, 158, 111 Destination: 333 Annotation: Subtype: Link Rect: 534, 420, 541, 432 Destination: 120 Annotation: Subtype: Link Rect: 384, 387, 391, 399 Destination: 333 Annotation: Subtype: Link Rect: 373, 321, 380, 333 Destination: 120 Annotation: Subtype: Link Rect: 538, 264, 550, 276 Destination: 333 Annotation: Subtype: Link Rect: 402, 209, 414, 221 Destination: 333 Annotation: Subtype: Link Rect: 489, 176, 496, 188 Destination: 141 Annotation: Subtype: Link Rect: 416, 77, 428, 89 Destination: 333 Thumb: true Page: Label: 6 Annotations: Annotation: Subtype: Link Rect: 266, 335, 349, 345 Annotation: Subtype: Link Rect: 130, 325, 208, 335 Annotation: Subtype: Link Rect: 407, 325, 508, 335 Annotation: Subtype: Link Rect: 90, 212, 103, 224 Destination: 333 Annotation: Subtype: Link Rect: 165, 113, 256, 125 Annotation: Subtype: Link Rect: 480, 289, 492, 301 Destination: 333 Annotation: Subtype: Link Rect: 490, 77, 497, 89 Destination: 141 Annotation: Subtype: Link Rect: 554, 44, 561, 56 Destination: 333 Thumb: true Page: Label: 7 Annotations: Annotation: Subtype: Link Rect: 155, 694, 162, 706 Destination: 141 Annotation: Subtype: Link Rect: 188, 694, 287, 706 Annotation: Subtype: Link Rect: 68, 617, 75, 629 Destination: 141 Annotation: Subtype: Link Rect: 256, 468, 263, 480 Destination: 333 Annotation: Subtype: Link Rect: 158, 292, 165, 304 Destination: 192 Annotation: Subtype: Link Rect: 45, 269, 296, 294 Annotation: Subtype: Link Rect: 91, 248, 98, 260 Destination: 192 Annotation: Subtype: Link Rect: 91, 237, 98, 249 Destination: 192 Annotation: Subtype: Link Rect: 258, 215, 265, 227 Destination: 192 Annotation: Subtype: Link Rect: 124, 193, 131, 205 Destination: 192 Annotation: Subtype: Link Rect: 217, 171, 224, 183 Destination: 192 Annotation: Subtype: Link Rect: 121, 116, 212, 128 Annotation: Subtype: Link Rect: 354, 716, 448, 728 Annotation: Subtype: Link Rect: 307, 627, 558, 652 Annotation: Subtype: Link Rect: 539, 617, 551, 629 Destination: 333 Annotation: Subtype: Link Rect: 308, 606, 320, 618 Destination: 333 Annotation: Subtype: Link Rect: 528, 529, 535, 541 Destination: 205 Annotation: Subtype: Link Rect: 326, 518, 418, 530 Annotation: Subtype: Link Rect: 451, 463, 463, 475 Destination: 403 Annotation: Subtype: Link Rect: 333, 419, 345, 431 Destination: 403 Annotation: Subtype: Link Rect: 404, 331, 411, 343 Destination: 205 Annotation: Subtype: Link Rect: 434, 331, 531, 343 Annotation: Subtype: Link Rect: 457, 287, 464, 299 Destination: 333 Annotation: Subtype: Link Rect: 447, 177, 459, 189 Destination: 403 Annotation: Subtype: Link Rect: 462, 177, 474, 189 Destination: 403 Annotation: Subtype: Link Rect: 375, 166, 382, 178 Destination: 205 Annotation: Subtype: Link Rect: 466, 89, 473, 101 Destination: 205 Annotation: Subtype: Link Rect: 499, 78, 507, 90 Destination: 205 Thumb: true Page: Label: 8 Annotations: Annotation: Subtype: Link Rect: 168, 95, 180, 107 Destination: 403 Annotation: Subtype: Link Rect: 466, 196, 559, 208 Annotation: Subtype: Link Rect: 428, 108, 519, 120 Thumb: true Page: Label: 9 Annotations: Annotation: Subtype: Link Rect: 74, 115, 157, 125 Thumb: true Page: Label: 10 Annotations: Annotation: Subtype: Link Rect: 78, 705, 85, 717 Destination: 244 Annotation: Subtype: Link Rect: 115, 683, 206, 695 Annotation: Subtype: Link Rect: 235, 672, 242, 684 Destination: 244 Annotation: Subtype: Link Rect: 107, 639, 114, 651 Destination: 244 Annotation: Subtype: Link Rect: 144, 556, 156, 568 Destination: 403 Annotation: Subtype: Link Rect: 223, 413, 235, 425 Destination: 333 Annotation: Subtype: Link Rect: 260, 391, 267, 403 Destination: 244 Annotation: Subtype: Link Rect: 269, 347, 282, 359 Destination: 333 Annotation: Subtype: Link Rect: 264, 314, 271, 326 Destination: 244 Annotation: Subtype: Link Rect: 256, 77, 269, 89 Destination: 403 Annotation: Subtype: Link Rect: 78, 44, 90, 56 Destination: 403 Annotation: Subtype: Link Rect: 426, 694, 433, 706 Destination: 256 Annotation: Subtype: Link Rect: 475, 639, 482, 651 Destination: 244 Annotation: Subtype: Link Rect: 428, 540, 435, 552 Destination: 256 Annotation: Subtype: Link Rect: 477, 496, 489, 508 Destination: 403 Annotation: Subtype: Link Rect: 427, 474, 439, 486 Destination: 403 Annotation: Subtype: Link Rect: 443, 474, 455, 486 Destination: 403 Annotation: Subtype: Link Rect: 507, 452, 515, 464 Destination: 244 Annotation: Subtype: Link Rect: 467, 430, 475, 442 Destination: 256 Annotation: Subtype: Link Rect: 433, 231, 445, 243 Destination: 403 Annotation: Subtype: Link Rect: 536, 187, 549, 199 Destination: 403 Annotation: Subtype: Link Rect: 378, 165, 390, 177 Destination: 403 Annotation: Subtype: Link Rect: 322, 110, 329, 122 Destination: 305 Annotation: Subtype: Link Rect: 331, 110, 344, 122 Destination: 333 Annotation: Subtype: Link Rect: 389, 88, 401, 100 Destination: 333 Thumb: true Page: Label: 11 Thumb: true Page: Label: 12 Annotations: Annotation: Subtype: Link Rect: 108, 179, 219, 189 Annotation: Subtype: Link Rect: 367, 110, 379, 122 Destination: 333 Annotation: Subtype: Link Rect: 382, 110, 394, 122 Destination: 403 Thumb: true Page: Label: 13 Annotations: Annotation: Subtype: Link Rect: 218, 705, 230, 717 Destination: 403 Annotation: Subtype: Link Rect: 233, 705, 245, 717 Destination: 403 Annotation: Subtype: Link Rect: 126, 584, 139, 596 Destination: 403 Annotation: Subtype: Link Rect: 50, 551, 62, 563 Destination: 403 Annotation: Subtype: Link Rect: 65, 551, 78, 563 Destination: 403 Annotation: Subtype: Link Rect: 268, 496, 280, 508 Destination: 403 Annotation: Subtype: Link Rect: 167, 452, 179, 464 Destination: 403 Annotation: Subtype: Link Rect: 117, 430, 129, 442 Destination: 403 Annotation: Subtype: Link Rect: 162, 408, 175, 420 Destination: 333 Annotation: Subtype: Link Rect: 128, 386, 141, 398 Destination: 403 Annotation: Subtype: Link Rect: 274, 375, 287, 387 Destination: 333 Annotation: Subtype: Link Rect: 103, 320, 115, 332 Destination: 333 Annotation: Subtype: Link Rect: 227, 232, 240, 244 Destination: 403 Annotation: Subtype: Link Rect: 204, 221, 216, 233 Destination: 403 Annotation: Subtype: Link Rect: 86, 188, 98, 200 Destination: 403 Annotation: Subtype: Link Rect: 206, 144, 219, 156 Destination: 403 Annotation: Subtype: Link Rect: 222, 144, 234, 156 Destination: 403 Annotation: Subtype: Link Rect: 247, 122, 260, 134 Destination: 403 Annotation: Subtype: Link Rect: 180, 78, 207, 90 Destination: 403 Annotation: Subtype: Link Rect: 50, 67, 62, 79 Destination: 403 Annotation: Subtype: Link Rect: 277, 56, 289, 68 Destination: 403 Annotation: Subtype: Link Rect: 436, 727, 448, 739 Destination: 403 Annotation: Subtype: Link Rect: 456, 628, 468, 640 Destination: 403 Annotation: Subtype: Link Rect: 334, 573, 346, 585 Destination: 403 Annotation: Subtype: Link Rect: 336, 540, 348, 552 Destination: 478 Annotation: Subtype: Link Rect: 539, 496, 551, 508 Destination: 478 Annotation: Subtype: Link Rect: 308, 485, 320, 497 Destination: 478 Annotation: Subtype: Link Rect: 373, 452, 385, 464 Destination: 478 Annotation: Subtype: Link Rect: 388, 452, 400, 464 Destination: 478 Annotation: Subtype: Link Rect: 311, 111, 323, 123 Destination: 333 Annotation: Subtype: Link Rect: 457, 89, 469, 101 Destination: 403 Thumb: true Page: Label: 14 Annotations: Annotation: Subtype: Link Rect: 158, 683, 171, 695 Destination: 478 Annotation: Subtype: Link Rect: 231, 573, 243, 585 Destination: 478 Annotation: Subtype: Link Rect: 116, 474, 128, 486 Destination: 403 Annotation: Subtype: Link Rect: 270, 452, 282, 464 Destination: 333 Annotation: Subtype: Link Rect: 57, 164, 307, 189 Annotation: Subtype: Link Rect: 319, 616, 401, 628 Annotation: Subtype: Link Rect: 482, 515, 531, 527 Annotation: Subtype: Link Rect: 423, 393, 523, 405 Annotation: Subtype: Link Rect: 331, 371, 415, 383 Annotation: Subtype: Link Rect: 322, 360, 425, 372 Annotation: Subtype: Link Rect: 323, 294, 332, 304 Destination: 39 Annotation: Subtype: Link Rect: 323, 264, 332, 274 Destination: 39 Annotation: Subtype: Link Rect: 332, 234, 492, 245 Annotation: Subtype: Link Rect: 323, 224, 332, 235 Destination: 39 Annotation: Subtype: Link Rect: 332, 204, 480, 215 Annotation: Subtype: Link Rect: 323, 194, 332, 205 Destination: 39 Annotation: Subtype: Link Rect: 398, 164, 553, 175 Annotation: Subtype: Link Rect: 323, 154, 332, 165 Destination: 39 Annotation: Subtype: Link Rect: 323, 124, 332, 135 Destination: 39 Annotation: Subtype: Link Rect: 332, 94, 478, 105 Annotation: Subtype: Link Rect: 323, 84, 332, 95 Destination: 39 Annotation: Subtype: Link Rect: 332, 45, 492, 55 Thumb: true Page: Label: 15 Annotations: Annotation: Subtype: Link Rect: 51, 728, 59, 739 Destination: 39 Annotation: Subtype: Link Rect: 60, 698, 218, 709 Annotation: Subtype: Link Rect: 51, 688, 59, 699 Destination: 39 Annotation: Subtype: Link Rect: 60, 659, 219, 669 Annotation: Subtype: Link Rect: 46, 649, 59, 659 Destination: 39 Annotation: Subtype: Link Rect: 60, 619, 219, 629 Annotation: Subtype: Link Rect: 46, 609, 59, 619 Destination: 39 Annotation: Subtype: Link Rect: 60, 569, 211, 580 Annotation: Subtype: Link Rect: 46, 559, 59, 570 Destination: 39 Annotation: Subtype: Link Rect: 124, 529, 274, 540 Annotation: Subtype: Link Rect: 46, 519, 59, 530 Destination: 39 Annotation: Subtype: Link Rect: 60, 479, 208, 490 Annotation: Subtype: Link Rect: 46, 469, 59, 480 Destination: 39 Annotation: Subtype: Link Rect: 148, 449, 283, 460 Annotation: Subtype: Link Rect: 46, 439, 59, 450 Destination: 39 Annotation: Subtype: Link Rect: 60, 409, 208, 420 Annotation: Subtype: Link Rect: 46, 400, 59, 410 Destination: 39 Annotation: Subtype: Link Rect: 60, 370, 209, 380 Annotation: Subtype: Link Rect: 46, 360, 59, 370 Destination: 39 Annotation: Subtype: Link Rect: 60, 330, 205, 340 Annotation: Subtype: Link Rect: 46, 320, 59, 331 Destination: 39 Annotation: Subtype: Link Rect: 117, 300, 258, 311 Annotation: Subtype: Link Rect: 46, 290, 59, 301 Destination: 39 Annotation: Subtype: Link Rect: 60, 260, 209, 271 Annotation: Subtype: Link Rect: 46, 250, 59, 261 Destination: 39 Annotation: Subtype: Link Rect: 60, 220, 201, 231 Annotation: Subtype: Link Rect: 46, 210, 59, 221 Destination: 39 Annotation: Subtype: Link Rect: 60, 190, 259, 201 Annotation: Subtype: Link Rect: 46, 180, 59, 191 Destination: 39 Annotation: Subtype: Link Rect: 113, 160, 250, 171 Annotation: Subtype: Link Rect: 46, 150, 59, 161 Destination: 39 Annotation: Subtype: Link Rect: 113, 121, 262, 131 Annotation: Subtype: Link Rect: 46, 111, 59, 121 Destination: 39 Annotation: Subtype: Link Rect: 126, 81, 266, 91 Annotation: Subtype: Link Rect: 46, 71, 59, 81 Destination: 39 Annotation: Subtype: Link Rect: 308, 708, 321, 719 Destination: 39 Annotation: Subtype: Link Rect: 321, 678, 456, 689 Annotation: Subtype: Link Rect: 308, 669, 321, 679 Destination: 39 Annotation: Subtype: Link Rect: 308, 639, 321, 649 Destination: 39 Annotation: Subtype: Link Rect: 308, 609, 321, 619 Destination: 39 Annotation: Subtype: Link Rect: 308, 579, 321, 590 Destination: 86 Annotation: Subtype: Link Rect: 321, 559, 483, 570 Annotation: Subtype: Link Rect: 308, 549, 321, 560 Destination: 86 Annotation: Subtype: Link Rect: 321, 509, 500, 520 Annotation: Subtype: Link Rect: 308, 499, 321, 510 Destination: 86 Annotation: Subtype: Link Rect: 390, 479, 542, 490 Annotation: Subtype: Link Rect: 308, 469, 321, 480 Destination: 86 Annotation: Subtype: Link Rect: 423, 449, 550, 460 Annotation: Subtype: Link Rect: 308, 439, 321, 450 Destination: 104 Annotation: Subtype: Link Rect: 321, 400, 480, 410 Annotation: Subtype: Link Rect: 308, 390, 321, 400 Destination: 104 Annotation: Subtype: Link Rect: 374, 360, 523, 370 Annotation: Subtype: Link Rect: 308, 350, 321, 360 Destination: 104 Annotation: Subtype: Link Rect: 321, 310, 459, 321 Annotation: Subtype: Link Rect: 308, 300, 321, 311 Destination: 104 Annotation: Subtype: Link Rect: 321, 270, 469, 281 Annotation: Subtype: Link Rect: 308, 260, 321, 271 Destination: 120 Annotation: Subtype: Link Rect: 308, 230, 321, 241 Destination: 120 Annotation: Subtype: Link Rect: 308, 210, 321, 221 Destination: 120 Annotation: Subtype: Link Rect: 321, 170, 496, 181 Annotation: Subtype: Link Rect: 308, 160, 321, 171 Destination: 141 Annotation: Subtype: Link Rect: 373, 121, 501, 131 Annotation: Subtype: Link Rect: 308, 111, 321, 121 Destination: 141 Annotation: Subtype: Link Rect: 308, 81, 321, 91 Destination: 158 Thumb: true Page: Label: 16 Annotations: Annotation: Subtype: Link Rect: 58, 728, 70, 739 Destination: 158 Annotation: Subtype: Link Rect: 71, 698, 247, 709 Annotation: Subtype: Link Rect: 58, 688, 70, 699 Destination: 158 Annotation: Subtype: Link Rect: 71, 659, 245, 669 Annotation: Subtype: Link Rect: 58, 649, 70, 659 Destination: 158 Annotation: Subtype: Link Rect: 71, 619, 242, 629 Annotation: Subtype: Link Rect: 58, 609, 70, 619 Destination: 158 Annotation: Subtype: Link Rect: 71, 569, 250, 580 Annotation: Subtype: Link Rect: 58, 559, 70, 570 Destination: 192 Annotation: Subtype: Link Rect: 141, 539, 303, 550 Annotation: Subtype: Link Rect: 58, 529, 70, 540 Destination: 213 Annotation: Subtype: Link Rect: 71, 479, 242, 490 Annotation: Subtype: Link Rect: 58, 469, 70, 480 Destination: 213 Annotation: Subtype: Link Rect: 71, 439, 189, 450 Annotation: Subtype: Link Rect: 58, 429, 70, 440 Destination: 213 Annotation: Subtype: Link Rect: 58, 400, 70, 410 Destination: 213 Annotation: Subtype: Link Rect: 152, 360, 300, 370 Annotation: Subtype: Link Rect: 58, 350, 70, 360 Destination: 213 Annotation: Subtype: Link Rect: 71, 330, 222, 340 Annotation: Subtype: Link Rect: 58, 320, 70, 331 Destination: 213 Annotation: Subtype: Link Rect: 71, 290, 214, 301 Annotation: Subtype: Link Rect: 58, 280, 70, 291 Destination: 213 Annotation: Subtype: Link Rect: 133, 260, 286, 271 Annotation: Subtype: Link Rect: 58, 250, 70, 261 Destination: 213 Annotation: Subtype: Link Rect: 71, 220, 189, 231 Annotation: Subtype: Link Rect: 58, 210, 70, 221 Destination: 213 Annotation: Subtype: Link Rect: 141, 190, 303, 201 Annotation: Subtype: Link Rect: 58, 180, 70, 191 Destination: 256 Annotation: Subtype: Link Rect: 124, 160, 281, 171 Annotation: Subtype: Link Rect: 58, 150, 70, 161 Destination: 268 Annotation: Subtype: Link Rect: 137, 121, 295, 131 Annotation: Subtype: Link Rect: 58, 111, 70, 121 Destination: 268 Annotation: Subtype: Link Rect: 71, 81, 229, 91 Annotation: Subtype: Link Rect: 58, 71, 70, 81 Destination: 268 Annotation: Subtype: Link Rect: 332, 718, 528, 729 Annotation: Subtype: Link Rect: 319, 708, 332, 719 Destination: 268 Annotation: Subtype: Link Rect: 332, 678, 480, 689 Annotation: Subtype: Link Rect: 319, 669, 332, 679 Destination: 268 Annotation: Subtype: Link Rect: 403, 649, 532, 659 Annotation: Subtype: Link Rect: 319, 639, 332, 649 Destination: 268 Annotation: Subtype: Link Rect: 434, 619, 563, 629 Annotation: Subtype: Link Rect: 319, 609, 332, 619 Destination: 268 Annotation: Subtype: Link Rect: 403, 589, 531, 599 Annotation: Subtype: Link Rect: 319, 579, 332, 590 Destination: 268 Annotation: Subtype: Link Rect: 396, 559, 522, 570 Annotation: Subtype: Link Rect: 319, 549, 332, 560 Destination: 268 Annotation: Subtype: Link Rect: 319, 519, 332, 530 Destination: 268 Annotation: Subtype: Link Rect: 332, 489, 484, 500 Annotation: Subtype: Link Rect: 319, 479, 332, 490 Destination: 268 Annotation: Subtype: Link Rect: 398, 449, 523, 460 Annotation: Subtype: Link Rect: 319, 439, 332, 450 Destination: 268 Annotation: Subtype: Link Rect: 332, 409, 457, 420 Annotation: Subtype: Link Rect: 319, 400, 332, 410 Destination: 268 Annotation: Subtype: Link Rect: 332, 370, 480, 380 Annotation: Subtype: Link Rect: 319, 360, 332, 370 Destination: 268 Annotation: Subtype: Link Rect: 332, 340, 487, 350 Annotation: Subtype: Link Rect: 319, 330, 332, 340 Destination: 268 Annotation: Subtype: Link Rect: 394, 300, 532, 311 Annotation: Subtype: Link Rect: 319, 290, 332, 301 Destination: 268 Annotation: Subtype: Link Rect: 332, 260, 507, 271 Annotation: Subtype: Link Rect: 319, 250, 332, 261 Destination: 268 Annotation: Subtype: Link Rect: 332, 220, 452, 231 Annotation: Subtype: Link Rect: 319, 210, 332, 221 Destination: 268 Annotation: Subtype: Link Rect: 319, 180, 332, 191 Destination: 268 Annotation: Subtype: Link Rect: 332, 150, 457, 161 Annotation: Subtype: Link Rect: 319, 140, 332, 151 Destination: 268 Annotation: Subtype: Link Rect: 332, 111, 470, 121 Annotation: Subtype: Link Rect: 319, 101, 332, 111 Destination: 268 Annotation: Subtype: Link Rect: 332, 61, 511, 71 Thumb: true Page: Label: 17 Annotations: Annotation: Subtype: Link Rect: 46, 728, 59, 739 Destination: 268 Annotation: Subtype: Link Rect: 60, 708, 190, 719 Annotation: Subtype: Link Rect: 46, 698, 59, 709 Destination: 268 Annotation: Subtype: Link Rect: 46, 669, 59, 679 Destination: 268 Annotation: Subtype: Link Rect: 60, 639, 176, 649 Annotation: Subtype: Link Rect: 46, 629, 59, 639 Destination: 268 Annotation: Subtype: Link Rect: 321, 718, 474, 729 Annotation: Subtype: Link Rect: 308, 708, 321, 719 Destination: 268 Annotation: Subtype: Link Rect: 391, 678, 542, 689 Annotation: Subtype: Link Rect: 308, 669, 321, 679 Destination: 305 Annotation: Subtype: Link Rect: 390, 649, 522, 659 Annotation: Subtype: Link Rect: 308, 639, 321, 649 Destination: 305 Annotation: Subtype: Link Rect: 321, 609, 494, 619 Annotation: Subtype: Link Rect: 365, 38, 490, 46 Annotation: Subtype: Link Rect: 446, 30, 495, 39 Annotation: Subtype: Link Rect: 46, 15, 129, 24 Thumb: true Checksum: 06261770 Type: CRC32 Checksum: d5fcbdd380a782432c307835bf5a372e Type: MD5 Checksum: 73658e8bf1e1580b722c555d967f8a648eea2dde Type: SHA-1 Checksum: dfe5882a44b3c63ba6b9e15b4e78a47ef75e295d752f0fd00d5eab19aafd47df Type: SHA-256