Sterilization study National Library of the Czech Republic
Materials, aids, instruments, equipments : Distilled water Cotton-wool tampons Substrate for growth of moulds (Czapek-Doxovs agar-CDA) Petri dishes Fume hood Laboratory tub Plastic bags Freezer (Zanussi ZCF 410) Vacuum-freeze drying chamber Ethylene oxide chamber (Matachana) Aseptic box (Fatran LF) Biological thermostat (BT 120 M) Optical microscope (Biolar) Autoclave (Tuttnauer 3870 M) Digital camera (Olympus C-990) Thermohygrometer (Testo 601) Mechanical measuring instruments on book dimensions It were found out general charakteristics of volumes (type of volume, material of bookbinding, material of book block) and were measured some physical property (dimension, temperature, relative humidity). Dimension of volumes (height, width, thickness) were measured before and after sterilization by the means mechanical measuring instruments on book dimensions. Temperature and relative humidity were measured inside of volumes before sterilization and after sterilization by the means termohygrometer Testo 601. Then were dilated by the means cloth chosen pages in the volumes to were full wet at wetting process. Wetting process (National Library of the Czech Republic) Dilated volumes were diped to the laboratory tub with distilled water and one day were left dip into distilled water. Then volumes were uncased from laboratory tub, shortly left standing to drain, and this volumes were given to the plastic bags, which we were airtightly closed on 7 days (simulate full-flood conditions). Freezing (National Library of the Czech Republic) Wet volumes were wraped to the plastic bags and were situated to the freezer. Freezing was perfomed in freezer at temperature -20oC. Vacuum-freeze drying (State Central Archives in Prague)
Frozen volumes were free situated in vacuum-freeze drying. Drying process was perfomed in vacuum-freeze drying chamber at temperature from -12 oC to -18oC and pressure 2 mbar. Drying process took about three weeks. Sterilization (State Central Archives in Prague) Sterilization process was performed in ethylene oxide chamber. Own sterilization process took one day. Then followed ventilation of volume in ventilation tunnel about one week. Microbiological inspection (National Library of the Czech Republic) Microbiological inspection carry out by dry cotton-wool tampons from bookbinding and book block of volume (10x10 cm) before and after sterilization. Then these tampons inoculates on sterile Petri dishes with Czapek- Doxovs agar (CDA) in the aseptic box. Cultivation of fungies last in the biological thermostat about 7-14 days at temperature 25oC. Then we carry out quantitative and qualitative identification of moulds. Quantitative identifications include number of colonies of moulds. Qualitative identifications include observe microscopic morphology of moulds and find out of species of moulds. Evaluation : Table 1 : Physical property of volumes Volume E-007 E-014 E-021 E-029 E-032 E-040 E-048 E-051 E-054 E-062 E-075 E-083 E-110 E-113 Type of volume Print Print Print Print Print Print Print Print Print Print Print Print Print Print Material of bookbinding Leather Leather Leather Leather Leather Leather Textile Textile Textile Leather Textile Leather Paper Textile Leather Paper Leather Material of book block Paper Paper Paper Paper Paper Paper Paper Paper Paper Paper Paper Paper Paper Paper Dimension /height x width x thickness/ (mm) 127x82x16 209x125x43 179x109x27 158x102x25 220x134x45 258x166x65 208x142x44 197x140x30 279x193x49 254x188x47 237x164x39 222x155x43 189x133x38 217x162x30 Temperature (oC) 20,4 20,3 20,3 20,4 20,7 20,6 20,6 20,7 20,4 19,9 20,9 21,0 20,8 19,8 Relative humidity (%) 25,7 23,6 24,8 25,8 26,3 23,3 24,5 25,4 23,5 22,7 22,6 22,5 24,3 23,7 Dimension /height x width x thickness/ (mm) 125x77x28 209x122x75 178x104x38 157x96x42 220x136x64 259x164x90 211x141x55 198x138x34 284x189x90 256x185x64 227x162x53 227x153x64 188x127x57 221x161x57 Temperature (oC) 21,3 21,1 21,4 21,1 21,5 20,9 21,3 21,0 21,1 21,2 21,0 21,1 21,2 21,0 Relative humidity (%) 36,7 32,7 35,4 32,9 32,7 43,3 33,5 34,5 38,9 37,9 33,7 28,6 34,6 35,2
E-121 E-124 E-127 E-130 E-138 E-150 E-153 E-170
Print Print Print Print Print Print Print Print
Leather Leather Textile Textile Leather Textile Textile Plastic
Paper Paper Paper Paper Paper Paper Paper Paper
237x163x48 210x140x27 247x169x29 244x168x40 212x158x31 243x162x36 247x170x29 260x201x27
19,4 20,5 20,1 19,0 20,4 20,3 20,5 21,0
23,5 24,3 22,6 22,3 23,4 23,9 22,3 24,6
238x164x66 208x136x36 249x169x46 246x170x61 210x157x45 245x159x57 250x169x48 261x200x51

21,1 21,0 21,2 21,4 21,3 21,4 21,3 20,9
33,0 35,8 38,8 37,0 35,3 35,7 35,4 34,1
Table 2 : Quantitative and qualitative identifications of moulds on volumes before and after sterilization Before sterilization Identificated moulds Identificated moulds (bookbinding) (book block) 6x Aspergillus niger 0 1x Penicillium chrysogenum 1x Aspergillus versicolor (total : 8) (total : 0) 3x Aspergillus niger 1x Aspergillus flavus 2x Penicillium chrysogenum 1x Aspergillus flavus 1x Aspergillus versicolor (total : 7) (total : 1) 6x Aspergillus niger 1x Aspergillus flavus 6x Penicillium chrysogenum 2x Aspergillus flavus (total : 14) (total : 1) 12x Aspergillus niger 0 (total : 12) (total : 0) 4x Penicillium chrysogenum 1x Penicillium chrysogenum 3x Aspergillus niger 1x Cladosporium cladosporoides 3x Aspergillus flavus 3x Trichoderma harzianum 1x Cladosporium cladosporoides (total : 14) (total : 2) 4x Penicillium chrysogenum 0 4x Aspergillus flavus After sterilization Identificated moulds Identificated moulds (bookbinding) (book block) 1x Aspergillus niger 0

Volume

Effect of Sterilization (bookbinding) 87,5%
Effect of Sterilization (book block)

(total : 1) 0

(total : 0) 0

(total : 0) 0 100% 100%

E-029 E-032
(total : 0) 3x Aspergillus niger (total : 3) 0
(total : 0) 0 (total : 0) 0

75% 100% 100%

1x Aspergillus niger (total : 9) 9x Aspergillus niger 4x Penicillium chrysogenum 1x Aspergillus flavus 1x Aspergillus versicolor (total : 15) 11x Aspergillus niger 5x Penicillium chrysogenum 1x Aspergillus flavus 1x Cladosporium cladosporoides (total : 18) 11x Penicillium chrysogenum 3x Cladosporium cladosporoides 2x Aspergillus niger 1x Aspergillus flavus (total : 17) 2x Aspergillus versicolor 1x Penicillium chrysogenum 1x Cladosporium cladosporoides (total : 4) 3x Penicillium chrysogenum 1x Aspergillus flavus 1x Aspergillus versicolor (total : 5) 2x Aspergillus flavus 2x Penicillium chrysogenum (total : 4) 9x Penicillium chrysogenum 1x Aspergillus versicolor (total : 10) 11x Aspergillus niger 4x Penicillium chrysogenum 1x Cladosporium cladosporoides (total : 16) 3x Aspergillus niger 3x Penicillim chrysogenum 2x Cladosporium herbarum
(total : 0) (total : 0) 1x Penicillium chrysogenum 1x Aspergillus niger

(total : 0) 0 93,3% 100%

(total : 1) 2x Aspergillus niger 1x Penicillium chrysogenum
(total : 0) 2x Penicillium chrysogenum

(total : 3) 0

(total : 2) 3x Penicillium chrysogenum
(total : 0) 0 (total : 0) 1x Penicillium chrysogenum (total : 1) 0
(total : 0) 0 (total : 0) 2x Penicillium chrysogenum (total : 2) 0

(total : 0) 0 (total : 0) 0 (total : 0) 0
(total : 0) 1x Cladosporium herbarum
1x Aspergillus versicolor (total :9) 5x Aspergillus niger 2x Penicillium chrysogenum 1x Aspergillus versicolor 1x Scopulariopsis brevicaulis (total : 9) 5x Aspergillus niger 3x Aspergillus flavus 1x Penicillium chrysogenum (total : 9) 8x Penicillium chrysogenum 7x Aspergillus niger
(total : 1) 1x Aspergillus versicolor
(total : 1) 1x Aspergillus flavus 1x Penicillium chrysogenum (total : 2) 6x Trichoderma harzianum 1x Penicillium chrysogenum 1x Aspergillus flavus (total : 8) 1x Penicillium chrysogenum
(total : 0) 1x Penicillium chrysogenum
(total : 15) 3x Aspergillus niger 3x Penicillium chrysogenum 3x Trichoderma harzianum 2x Aspergillus flavus (total : 11) (total : 1) 12x Aspergillus niger 1x Penicillium chrysogenum 3x Penicillium chrysogenum 1x Scopulariopsis brevicaulis (total : 16) (total : 1) 11x Penicillium chrysogenum 3xPenicillium chrysogenum 3x Cladosporium cladosporoides 1xAspergillus versicolor 1x Aspergillus niger 1x Cladosporium cladosporoides (total : 15) (total : 5) 4x Aspergillus niger 0 2x Penicillium chrysogenum 2x Cladosporium cladosporoides 1x Aspergillus versicolor (total : 9) (total : 0)
(total : 2) 1x Penicillium chrysogenum 1x Aspergillus flavus
(total : 2) 3x Aspergillus niger
(total : 3) 1x Penicillium chrysogenum

(total : 0)

(total : 0) 93,2% 99,1%

Average

Number of colonies of moulds on volumes express low level of fungal pollution. Identificated moulds are usual species of moulds which are situated on book materials. Effect of sterilization on moulds which are situated on bookbinding is 93,2%. Effect of sterilization on moulds which are situated on book block is 99,1%. Identificated of moulds :
Aspergillus flavus Colonies are low and velvet or grainy. Surface of colonies is at first white later becomes yellow, yellow-green, yellow-brown or brown. Reverse is yellow (from pigment) or brown. Facilities of range : land, plant residues, foods (seeds), paper, leather, textile. It produce mycotoxins (aflatoxins ). Aspergillus niger Colonies are high and wooly. Surface of colonies is at first white later becomes brown or black. Reverse is colourless. Facilities of range : land, atmosphere, plant residues, paper, leather, textile. It produce citric acid and mycotoxins (malformin ). Aspergillus versicolor Colonies are low and velvet. Surface of colonies is at first white later becomes yellow, umber, yellow-brown, yellow-green or brown. Reverse is yellow, umber, red-brown (from pigment) or brown. Facilities of range : land, atmosphere, plant residues, foods, corn, paper, leather, textile. It produce mycotoxins (sterigmatocystin ). Cladosporium cladosporoides Colonies are low and velvet. Surface of colonies is brown. Reverse is brown-black. Facilities of range : atmosphere, plant residues, wet walls, paper, leather, textile. Cladosporium herbarum Colonies are low and felt or velvet. Surface of colonies is green-brown or black-brown. Reverse is black-gray or black. Facilities of range : plant residues, wet walls, paper, leather, textile. It produce mycotoxins (gibberelin ). Penicillium chrysogenum Colonies are low and velvet. Surface of colonies is at first white later becomes yellow-green, brown-green or blue-green with white hem. Reverse is yellow (from pigment), yellow-green or yellow-brown. Facilities of range : land, atmosphere, plant residues, foods, wet walls, paper, leather, textile. It produce antibiotic penicillin and mycotoxins (ochratoxin ). Scopulariopsis brevicaulis Colonies are low and velvet or wooly. Surface of colonies is at first white later becomes cinnamon-brown or yellow-brown. Reverse is colourless or light brown. Facilities of range : land, plant, paper, leather, textile. Trichoderma harzianum Colonies are low and wooly. Surface of colonies is at first white later becomes green. Reverse is colourless. Facilities of range : land, plant residues, foods, paper, leather, textile. It produce mycotoxins (chrysophanol ).