\r\n\tManagement of these disorders requires good clinical evaluation, diagnostic tests, appropriate therapy and huge healthcare cost. Sometimes multiple specialties (gastroenterologists, \r\n\tgastrointestinal motility specialists, otolaryngologists, surgeons, speech therapists, medical oncologists and radiation oncologists) are involved in the management of dysphonia and dysphagia. In the recent years, there have been many updates in the management of these disorders. This book will discuss systematically the different etiologies and management of dysphonia, maxillofacial, oropharyngeal and esophageal dysphagia. This book will be a good \r\n\tguide to the practicing physicians for the management of voice and swallowing disorders.
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1. Introduction
Epinephelus groupers (Perciformes, Serranidae) are widely distributed in tropical and subtropical waters [1] and comprise 89 species (valid names) in marine habitats worldwide [2]. Most known grouper species are in the Indian-Pacific Ocean, 11 species along the West Atlantic coast, nine species in the East Atlantic Ocean and Mediterranean, and eight species in the eastern Pacific Ocean. Only a few groupers are distributed across different oceans [1]. Forty-one species of groupers in total were found in coastal waters of Taiwan [3].
Grouper is an important aquatic product in the world. In addition to abundant grouper caught at sea, the artificial breeding grouper is also a major aquatic product in the fishery trade. In the past, most grouper fry were from Southeast Asian countries such as the Philippines, Indonesia, and Thailand. However, survival rates markedly decreased due to catching and transportation. Nowadays, breeding techniques have been completely established for major commercial groupers, and so most grouper fry are bought from artificial breeding farms. Currently, Epinephelus akaara, E. areolatus, E. awoara, E. bleekeri, E. bruneus, E. fuscoguttatus, E. lanceolatus, E. septemfasciatus, E. tauvina, E. coioides, and E. malabaricus can be artificially reared and bred, especially E. malabaricus, which is the most successful case. Groupers have similar external morphologies, and their body color characteristics are not stable. Juveniles and adult fishes may show completely different color patterns. Therefore, it is often impossible to effectively distinguish species with similar morphologies in the adult stage [1, 4, 5]. As to their mating systems, incorrect identification of parents and progeny in rearing and breeding farms may cause artificial full-breeding plans and hybridization strategies to fail; moreover, this will result in significant fishery losses [1, 5, 6].
Traditionally, grouper species were classified using morphological and skeletal features [1, 7, 8, 9]. In the past two decades, molecular genetic technology has been dramatically developed and is now widely used in taxonomic and systematics studies. As Ref. [5] analyzed 42 species of grouper including three genera (Epinephelus, Cephalopholis, and Mycteroperca) using partial 16S ribosomal (r)DNA sequences. Results of that phylogenetic study revealed that both genera Epinephelus and Mycteroperca belong to the same clade, and it was inferred that Serranidae comprised a paraphyletic group.
Nowadays ichthyologists also use variable staining methods to obtain cytogenetic information of fish [10, 11]. According to previous studies, the number of chromosomes in groupers are 2n = 48, most of which are telocentric chromosomes, and fundamental numbers range 48–62 [12]. Some reports on the cytogenetics of grouper indicated that silver-binding nucleolar organizing regions (Ag-NORs) are highly conserved on the chromosome 24, but variations occur in the location between different groupers [13, 14, 15, 16, 17, 18]. It is generally believed that such variations may be caused by an inversion of the arms during chromosome evolution. To study an evolutionary model of chromosomes and identify species, staining techniques were used often to analyze the karyotype and cytogenetics of groupers.
More than ten groupers have been successfully cultivated in Taiwan. However, most groupers have similar external morphologies, and their color patterns are quite unstable. Often grouper in different life stages exhibit inconsistent color distributions that resulted in the species identification of grouper fry being controversial or confusing [1, 5]. In the aquaculture industry, misidentification frequently occurs in different growth stages of groupers, and this can cause serious problems, such as chaos of market prices, interspecific ecological competition, and breeding strategy failures.
It is important to understand the karyotype and phylogeny of cultured grouper for a successful strategy of genetic breeding. That is when studying hybridization strategies of groupers, selecting similar karyotypes and closely related species for the parents may result in relatively higher success potential for hybridization. Therefore, the establishment of grouper karyotype and barcode data in this study will provide more-perfect genetic bases for species identification to improve possibilities for genetic breeding. The present study analyzed the mitochondrial cytochrome (Cyt) b gene sequences and chromosomal characters of seven cultured groupers in Taiwan. These results will provide farmers with more genetic information of groupers to develop useful breeding strategies for hybridization in the future.
2. Materials and methods
2.1. Sampling
Seven groupers, Epinephaluslanceolatus, E. tukula, E. flavocaeruleus, E. polyphekadion, E. fuscoguttatus, E. coioides, and Plectropomusleopardus, were collected from fish markets in Tungkang, southern Taiwan (Figure 1) for chromosome preparation and DNA sequence analysis. A piece of muscle tissue from each specimen was preserved in 95% ethanol (EtOH) and stored at the Fish Biology Lab in National Pingtung University of Science and Technology. Seven species were used for the karyotype analysis and Cyt b gene sequencing.
Figure 1.
Sampling location of groupers.
2.2. Chromosomal preparation and karyotype analyses of groupers
The cell culture solution contained Eagle’s minimal essential medium (MEM) with 15% fetal bovine serum and 0.0001% colchicine, followed by filter-sterilization (0.45 μm). Kidney tissue was cut and placed in the cell culture solution. The solution tubes were placed on a rotary shaker (100 rpm) and then incubated at room temperature for 2 h to allow cells to remain in the metaphase of the cell cycle. The cell culture solution was centrifuged at 3000 rpm for 5 min, and the supernatant was discarded. KCl (at 0.075 M) was added and allowed to sit at room temperature for 30 min. After centrifugation at 3000 rpm for 5 min, the supernatant was discarded, and a freshly prepared fixative solution (methanol: acetic acid = 3:1) was added at room temperature for 15 min. The mixture was centrifuged at 3000 rpm for 5 min, the supernatant was discarded, and this step was repeated two or three times. The cell suspension was dropped onto a heated glass slide and air-dried. After the slide had been stained with 5% Giemsa dye for 10 min, it was rinsed with water and air-dried. The slide was mounted and observed by microscopy.
In addition, some fresh chromosome slides were stained with AgNO3. Two drops of 2% (w/v) gelatin and four drops of a 50% AgNO3 solution were mixed and then dropped onto a slide with a cover glass. These slides were incubated at 70°C until they presented a yellowish-brown color. The slides were gently rinsed with double-distilled (dd)H2O. After being air-dried at room temperature, the slides were mounted with gum arabic [19]. Chromosomes were observed with an optical microscope (Leica Microsystems, Wetzlar, Germany) (at 1000× with an oil lens). Digital images of the chromosomes were recorded and analyzed with a chromosome band analytical system (BandView 5.5, Applied Spectral Imaging, Migdal HaEmek, Israel). Chromosomes stained with Giemsa were classified into four groups, metacentric (m), submetacentric (sm), subtelocentric (st), and telocentric (t), according to the system described by [20]. Locations of chromosomes determined by AgNO3 staining were observed and marked on photos.
2.3. DNA isolation
Approximately 100 mg of muscle tissue from each specimen was put into an Eppendorf tube. Before DNA purification, the tube was placed in a 60°C oven for 10 min to evaporate the EtOH. Genomic DNA was isolated using a Gentra Puregene Core kit A (Qiagen, Venlo, the Netherlands), and the purified DNA specimen was dissolved in TE buffer (1 M Tris–HCl at pH 8.0 and 0.2 mL EDTA, 0.5 M). DNA concentrations were estimated using a Nanodrop 2000C spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) at an absorbance of 260 nm. The purity of DNA preparations was checked by the ratio of absorbances at 260 and 280 nm (A260/A280 ≥ 1.8). DNA stock solutions were stored in a − 20°C freezer.
2.4. Cyt b gene sequencing and analysis
In total, 50 μL of reactant of a polymerase chain reaction (PCR) contained 5 ng genomic DNA, 10 pmol each of the forward and reverse primers, 4 μL 2.5 mM dNTP, 0.2 μL 25 mM MgCl2, 1 U Taq polymerase, and 5 μL 10× buffer, with ddH2O added to 50 μL. The forward and reverse primers of the Cyt b gene were FOR (5’-CGAACGTTGATATGAAAAACCATCGTTG-3′) and UnvH (5’-ATCTTCGGTTTACAAGAC CGGTG-3′), respectively [6]. The Cyt b gene was amplified using a PCR machine (BIO-RAD MJ Mini Gradient Thermal Cycler, Conmall Biotechnology, Singapore) with initial denaturation at 95°C for 3 min; 35 cycles of 95°C for 1 min, 50°C for 1 min, and 72°C for 1 min; with a final extension of 72°C for 10 min. The reaction was cooled down to 25°C for 10 min. PCR products of the Cyt b gene were checked using 1% agarose gel electrophoresis and then stained with ethidium bromide (EtBr; 0.5 mg/mL). Target DNA fragments were eluted with a DNA Clean/Extraction kit (GeneMark, Taichung, Taiwan). Sizes of the purified DNA fragments were checked and then stored in a −20°C freezer. DNA fragments were directly sequenced on an Applied Biosystems (ABI, Foster City, CA, USA) automated ABI3730x1 DNA sequencer using a Bigdye sequencing kit (Perkin-Elmer, Wellesley, MA, USA). FOR or UnvH primers were used in the sequencing reaction, and the PCR cycle parameters for sequencing were 35 cycles of 30 s at 95°C, 30 s at 50°C, and 1 min at 72°C.
In total, seven Cyt b sequences were obtained in this study. Homologous sequences were aligned using ClustalW [21] and then manually checked. Interspecific genetic distances were analyzed using the Kimura-2-parameter (K2P) model [22], and numbers of different nucleotides were calculated with MEGA software [23]. The best-fitting models of DNA substitution were determined using the lowest Bayesian Information Criterion (BIC) scores [24]. The phylogenetic trees of Cyt b sequences were constructed using the Neighbor-joining (NJ) [25] and Maximum-likelihood (ML) methods [26]. Cluster confidence levels of Cyt b were assessed using a bootstrap analysis with 1000 replications [27].
3. Results
3.1. Karyotype analyses
In cytogenetic studies, Giemsa staining of seven groupers indicated that the diploid number of these species was 2n = 48. The karyotypic formulae were 2 sm + 46 t for E. coioides, E. fuscoguttatus, and E. tukula; 6 sm + 4 st + 38 t for E. lanceolatus; 2 st + 46 t for E. flavocaeruleus; 6 sm + 42 t for E. polyphekadion; and 48 t for P. leopardus. All of those specimens had a high number of telocentric chromosomes (38–48) and fundamental arm numbers (FNs) that ranged 48–54 (Figure 2, Table 1).
Figure 2.
Karyotype analyses of seven groupers: (a) Epinephelus coioides; (b) E. flavocaeruleus; (c) E. fuscoguttatus; (d) E. lanceolatus; (e) E. polyphekadion; (f) E. tukula; and (g) Plectropomus leopardus.
E., Epinephelus; P., Plectropomus; 2n, diploid number; FN, fundamental number; metacentric (m), submetacentric (sm), subtelocentric (st), and telocentric (t), according to the system described in Ref. [20].
In Ag-NO3 staining, four Epinephelus species (E. coioides, E. fuscoguttatus, E. tukula, and E. lanceolatus) and P. leopardus were completed. Epinephelus coioides, E. fuscoguttatus, and E. tukula had one pair of Ag-NORs located on the short arm of the sm chromosome; E. lanceolatus had two pairs of Ag-NORs located on the short arm of the sm chromosome; and P. leopardus had one pair of Ag-NORs, located near the centromere of larger telocentric chromosomes (Figure 3).
Figure 3.
Silver-binding nucleolar organizing regions (Ag-NORs) results from five groupers: (a) Epinephelus coioides; (b) E. fuscoguttatus; (c) E. tukula; (d) E. lanceolatus; and (e) Plectropomus leopardus. The arrows indicate Ag-NORs. The bar equals 5 μm.
3.2. Cyt b sequence analysis
All Cyt b gene sequences from seven groupers were 1141 bp for E. lanceolatus, E. tukula, E. flavocaeruleus, E. polyphekadion, E. fuscoguttatus, E. coioides, and P. leopardus. Percentages of nucleotide compositions did not significantly differ among these Epinephelus species, as the A + T ratios were in the range of 52.1% (E. flavocaeruleus) - 56.7% (E. polyphekadion). Interspecific p-distances and K2P genetic distances ranged 0.1149 and 0.1284 (E. tukula vs. E. coioides) to 0.1814 and 0.2138 (E. flavocaeruleus vs. E. polyphekadion) (Table 2). The best model of nucleotide evolution was estimated to be the TN93 + G + I model with BIC = 9065.099. The NJ and ML analyses showed that E. tukula and E. coioides had a close phylogenetic relationship with extremely high bootstrap support (Figure 4). This result agreed with the hypothesis that Epinephelus is a monophyletic group.
Code
Species name
1
2
3
4
5
6
7
1
E. lanceolatus
—
0.1422
0.1649
0.1474
0.1430
0.1333
0.2344
2
E. tukula
0.1635
—
0.1658
0.1684
0.1360
0.1149
0.2186
3
E. flavocaeruleus
0.1908
0.1934
—
0.1814
0.1578
0.1604
0.2272
4
E. polyphekadion
0.1690
0.1994
0.2138
—
0.1516
0.1595
0.2237
5
E. fuscoguttatus
0.1638
0.1561
0.1817
0.1751
—
0.1350
0.2123
6
E. coioides
0.1510
0.1284
0.1853
0.1858
0.1436
—
0.2307
7
P. leopardus
0.2859
0.2617
0.2738
0.2699
0.2529
0.2803
—
Table 2.
p-distance genetic distances (above the diagonal) and Kimura 2-parameter distances (below the diagonal) of cytochrome b gene sequences among Epinephelus groupers and the outgroup Plectropomus leopardus.
Figure 4.
(a) The Neighbor-joining and (b) the Maximum-likelihood trees among Epinephelus species based on the cytochrome b gene analysis.
4. Discussion
In this study, Epinephelinae fish (E. lanceolatus, E. tukula, E. flavocaeruleus, E. polyphekadion, E. fuscoguttatus, E. coioides, and P. leopardus) showed a common synapomorphic character of chromosomal number, 2n = 48, and high numbers of telocentric chromosomes (38–48). By sorting out the cytogenetic information of 23 Epinephelinae species, it was found that chromosomal numbers of these groupers were 48, showing highly conserved characteristics, and FNs ranged 48–62, with more than half of these groupers exhibiting FN = 48 characteristics (Table 1), in accordance with conservative chromosomal morphological features described in Ref. [28]. In the other hand, variations in FNs are mainly caused by chromosomal rearrangements and play important roles in the speciation process [29].
In cytogenetic studies, karyotypes, FNs, Ag-NORs, and C-bands were demonstrated to have interspecific specificities, and many studies used these techniques to explore interspecific evolutionary relationships [30, 31, 32]. Currently, reports related to chromosomes of Epinephelus groupers worldwide are only available for 23 of 89 groupers; e.g., karyotypes of E. marginatus were analyzed from three different sampling sites in the Mediterranean. Results showed chromosomal numbers of 2n = 48; conserved C-bands and Ag-NOR positions were observed on the 24th pair of chromosomes of specimens from all three samples, but those were also found on 2nd pair chromosomes of one specimen [18]. In order to confirm the above results, fluorescence in situ hybridization (FISH) was performed using 18S rDNA as a probe. Fluorescence reacted to the 2nd and 24th pairs of chromosomes confirming that a difference existed between samples. The authors reasoned that this may have been a species-specific manifestation, and further studies are required to confirm whether they can be population-specific markers.
Molecular phylogenetic analyses showed that both Plectropomus and Cephalopholis are more primitive genera than Epinephelus [5, 6, 12, 33, 34]. In this study, the chromosomal number of P. leopardus was 2n = 48 t. All current cytogenetic studies of Epinephelus groupers have shown that few of them are not composed of 2n = 48 t. These results support 48 t being an ancestral character of Serranidae fish [12], and Epinephelus groupers may be a later-derived genus.
In Ref. [12] observed three types of Ag-NORs distribution pattern: type I has only one pair of Ag-NORs located in the subcentromeric region of the acrocentric (t) chromosome, e.g., E. guaza, E. alexandrinus, E. caninus, E. fasciatomaculatus, E. fasciatus, and E. awoara; type II has one pair of Ag-NORs located in the subcentromeric region of the t chromosome pair and an extra pair of smaller Ag-NORs located on another pair of chromosomes, as in E. adscensionis, E. marginatus, and E. malabaricus; and type III has only one pair of Ag-NORs located on the short arm of bi-armed chromosomes, e.g., E. guttatus and E. coioides. Thus, based on the available cytogenetic data on the genus Epinephelus, most of the NORs of groupers are located on the 24th pair of chromosome (type I), and these results are consistent with those of [18]. In this study, E. fuscoguttatus, E. tukula, and E. lanceolatus also belonged to type III. It is generally believed that the appearance of one pair of Ag-NORs is the ancestral character of Serranidae fish [28]. However, when Ref. [12] classified this character and compared it to data of molecular phylogenies, results were found to be irrelevant. The authors believe that the contradiction between cytogenetic and molecular phylogenetic analyses may merely be the result of insufficient data.
Hybrid breeding often produces heterosis offspring, such as offspring with a fast growth rate, strong disease resistance, or diverse morphology. For example, Liu et al. crossed different carps to obtain hybrids with a high growth rate [35]. However, many studies have found that the success possibility and whether the offspring are fertile are related to the parental karyotypes. The parents having more-similar karyotypes can increase the success ratio of hybridization [36]. At present, completely cultured groupers mainly consist of E. akaara, E. areolatus, E. awoara, E. bleekeri, E. bruneus, E. fuscoguttatus, E. lanceolatus, E. septemfasciatus, E. tauvina, E. coioides, and E. malabaricus. Establishment of karyotypic data of these groupers can provide references for crossing strategies on farms. The genetic relationship and chromosome composition of hybrid progeny can also be confirmed by a karyotype test.
Species names of different groupers have always been confusing. Most groupers living coral reef areas have similar external morphologies, and their color characteristics also may change along with their living environment. Some larvae and juveniles may even have completely different color distributions from adults, such as E. lanceolatus which has three irregular black spots and a brilliant color as juveniles, but becomes dark brown as adults. Therefore, identifying groupers is often controversial [1, 4, 5]. For example, E. coioides and E. tauvina are very similar and difficult to distinguish in Taiwanese waters [37]. There is still much dispute over the taxonomy of groupers when using traditional morphology. Cyt b gene marker is of great help in identifying similar groupers or unidentifiable fry. In the future, this marker can also be used in aquaculture breeding to reduce failures and losses with artificial reproduction.
In this study, the results showed that different groupers can be identified by analyzing the Cyt b gene. The phylogenetic tree constructed from the Cyt b gene can distinguish Epinephelus groupers from those in the genus Plectropomus. However, groupers evolved as monophyletic group, the genus Plectropomus is a relatively primitive group in Epinephelinae.
Epinephelus lanceolatus was previously classified in the genus Promicrops by [38, 39], but [6] used Cyt b to study molecular phylogenetic relationships of six out of 28 genera in the Serranidae, suggested that Promicrops lanceolatus should be classified into Epinephelus. Phylogenetic trees constructed with the NJ and ML methods also revealed that E. lanceolatus has a close relationship with other Epinephelus groupers [6]. In addition, scientific names of seven farmed groupers have been identified to reduce confusion and controversy.
5. Conclusions
All chromosome numbers from seven groupers (Plectropomus leopardus, Epinephelus coioides, E. flavocaeruleus, E. fuscoguttatus, E. lanceolatus, E. polyphekadion, and E. Tukula) showed a common synapomorphic character of chromosomal number, 2n = 48. Four groupers, E. coioides, E. polyphekadion, E. fuscoguttatus, and E. tukula shared the same karyotype formula of 2 sm + 46 t. E. coioides, E. fuscoguttatus, and E. tukula had one pair of Ag-NORs located on the short arm of the sm chromosome. The mitochondrial Cyt b gene was used to analyze phylogenetic relationships among these species. We discovered that Epinephelus groupers should be classified as monophyly. The minimum genetic distance expressed between E. coioides and E. tukula was 0.1276. Results showed that E. coioides and E. tukula have similar genetic characters and cell karyotypes, and should be foremost considered for artificial hybridization strategies. Information on karyotypes of species within the Epinephelus is still insufficient, and further elucidation of karyotypes of Epinephelus will be a great help to future genetic breeding research.
Acknowledgments
The authors express their gratitude to IW Shih for assistance with laboratory work, and also thank Professor TB Yen for his help on image process and data integration.
Research funding was provided to MC Tseng by the Ministry of Science and Technology, Taiwan (NSC 102-2313-B-020-002).
Conflict of interest
Both authors, Mei-Chen Tseng and Kuan-Wei Shih declare that they have no conflict of interest.
\n',keywords:"barcode, cytochrome b, cytogenetic, genetic distance, hybridization",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/63181.pdf",chapterXML:"https://mts.intechopen.com/source/xml/63181.xml",downloadPdfUrl:"/chapter/pdf-download/63181",previewPdfUrl:"/chapter/pdf-preview/63181",totalDownloads:228,totalViews:190,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,dateSubmitted:"April 24th 2018",dateReviewed:"July 20th 2018",datePrePublished:"November 5th 2018",datePublished:null,readingETA:"0",abstract:"In this study, karyotypes and Cyt b gene sequences of seven different species of grouper including Plectropomus leopardus, Epinephelus coioides, E. flavocaeruleus, E. fuscoguttatus, E. lanceolatus, E. polyphekadion, and E. tukula were examined. All chromosome numbers from seven groupers were 2n = 48 with a high number of telocentric chromosomes (38–48) and fundamental arm numbers (FNs) (48–54). The mitochondrial Cyt b gene was used to establish the barcodes of seven groupers and analyze phylogenetic relationships among these species. We discovered that Epinephelus groupers should be classified as monophyly. The minimum genetic distance expressed between E. coioides and E. tukula was 0.1276. From results of the cytogenetic and molecular analyses, it was demonstrated that Plectropomus is a relatively primitive genus of grouper, while Epinephelus is a more-modern derived genus. Results also showed that E. coioides and E. tukula have similar genetic characters and karyotypes, and should be foremost considered for artificial hybridization strategies. Furthermore, information on karyotypes of species within the Epinephelus is still insufficient, and further elucidation of karyotypes of Epinephelus will be a great help to future genetic breeding research.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/63181",risUrl:"/chapter/ris/63181",book:{slug:"cytogenetics-past-present-and-further-perspectives"},signatures:"Mei-Chen Tseng and Kuan-Wei Shih",authors:[{id:"96064",title:"Prof.",name:"Mei-Chen",middleName:null,surname:"Tseng",fullName:"Mei-Chen Tseng",slug:"mei-chen-tseng",email:"mctseng@mail.npust.edu.tw",position:null,institution:{name:"National Pingtung University of Science and Technology",institutionURL:null,country:{name:"Taiwan"}}}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Materials and methods",level:"1"},{id:"sec_2_2",title:"2.1. Sampling",level:"2"},{id:"sec_3_2",title:"2.2. Chromosomal preparation and karyotype analyses of groupers",level:"2"},{id:"sec_4_2",title:"2.3. DNA isolation",level:"2"},{id:"sec_5_2",title:"2.4. Cyt b gene sequencing and analysis",level:"2"},{id:"sec_7",title:"3. Results",level:"1"},{id:"sec_7_2",title:"3.1. Karyotype analyses",level:"2"},{id:"sec_8_2",title:"3.2. Cyt b sequence analysis",level:"2"},{id:"sec_10",title:"4. Discussion",level:"1"},{id:"sec_11",title:"5. Conclusions",level:"1"},{id:"sec_12",title:"Acknowledgments",level:"1"},{id:"sec_12",title:"Conflict of interest",level:"1"}],chapterReferences:[{id:"B1",body:'Heemstra PC, Randall JE. Groupers of the world (Family Serranidae, subfamily Epinephelinae). An annotated and illustrated catalogue of the grouper, rockcod, hind, coral grouper and lyretail species known to date. FAO Fisheries Synopsis. 1993;125:1-382'},{id:"B2",body:'Wo RMS. Epinephelus. In: Bailly N, editor. Fish Base. World Register of Marine Species. [Accessed: 4 Apr 2012]. http://www.marinespecies.org/aphia.php?p=taxdetails&id=126068'},{id:"B3",body:'Shao KT. Taiwan Fish Database. 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Department of Aquaculture, National Pingtung University of Science and Technology, Pingtung, Taiwan, R.O.C.
Department of Aquaculture, National Pingtung University of Science and Technology, Pingtung, Taiwan, R.O.C.
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York and William Neal Reynolds",authors:[{id:"13418",title:"Prof.",name:"David",middleName:null,surname:"Jordan",fullName:"David Jordan",slug:"david-jordan"}]},{id:"13135",title:"Computational Biology, Protein Engineering, and Biosensor Technology: a Close Cooperation for Herbicides Monitoring",slug:"computational-biology-protein-engineering-and-biosensor-technology-a-close-cooperation-for-herbicide",signatures:"Giuseppina Rea, Fabio Polticelli, Amina Antonacci, Maya Lambreva, Sandro Pastorelli, Viviana Scognamiglio, Veranika Zobnina and Maria Teresa Giardi",authors:[{id:"13746",title:"Dr.",name:"Giuseppina",middleName:null,surname:"Rea",fullName:"Giuseppina Rea",slug:"giuseppina-rea"},{id:"15034",title:"Dr.",name:"Fabio",middleName:null,surname:"Polticelli",fullName:"Fabio Polticelli",slug:"fabio-polticelli"},{id:"15038",title:"Dr.",name:"Amina",middleName:null,surname:"Antonacci",fullName:"Amina Antonacci",slug:"amina-antonacci"},{id:"15039",title:"Dr.",name:"Sandro",middleName:null,surname:"Pastorelli",fullName:"Sandro Pastorelli",slug:"sandro-pastorelli"},{id:"15040",title:"Dr.",name:"Maya",middleName:null,surname:"Lambreva",fullName:"Maya Lambreva",slug:"maya-lambreva"},{id:"15041",title:"Dr.",name:"Maria Teresa",middleName:null,surname:"Giardi",fullName:"Maria Teresa Giardi",slug:"maria-teresa-giardi"},{id:"15178",title:"Dr.",name:"Veranika",middleName:null,surname:"Zobnina",fullName:"Veranika Zobnina",slug:"veranika-zobnina"},{id:"22620",title:"Dr.",name:"Viviana",middleName:null,surname:"Scognamiglio",fullName:"Viviana Scognamiglio",slug:"viviana-scognamiglio"}]},{id:"13136",title:"Statistical Based Real-Time Selective Herbicide Weed Classifier",slug:"statistical-based-real-time-selective-herbicide-weed-classifier",signatures:"Irshad Ahmad and Abdul Muhamin Naeem",authors:[{id:"13787",title:"Prof.",name:"Irshad",middleName:null,surname:"Ahmad",fullName:"Irshad Ahmad",slug:"irshad-ahmad"},{id:"23691",title:"Prof.",name:"Abdul Muhamin",middleName:null,surname:"Naeem",fullName:"Abdul Muhamin Naeem",slug:"abdul-muhamin-naeem"}]},{id:"13137",title:"Variable Rate Herbicide Application Using GPS and Generating a Digital Management Map",slug:"variable-rate-herbicide-application-using-gps-and-generating-a-digital-management-map",signatures:"Majid Rashidi and Davood Mohammadzamani",authors:[{id:"14094",title:"Prof.",name:"Majid",middleName:null,surname:"Rashidi",fullName:"Majid Rashidi",slug:"majid-rashidi"}]},{id:"13138",title:"Soil Electrical Conductivity as One Possible Tool for Predicting of Cirsium Arvense Infestation Occurrence",slug:"soil-electrical-conductivity-as-one-possible-tool-for-predicting-of-cirsium-arvense-infestation-occu",signatures:"Milan Kroulik, Atonin Slejska, Dana Kokoskova and Veronika Venclova",authors:[{id:"13477",title:"Dr.",name:"Milan",middleName:null,surname:"Kroulik",fullName:"Milan Kroulik",slug:"milan-kroulik"},{id:"14742",title:"Prof.",name:"Antonin",middleName:null,surname:"Slejska",fullName:"Antonin Slejska",slug:"antonin-slejska"},{id:"14743",title:"Dr.",name:"Dana",middleName:null,surname:"Kokoskova",fullName:"Dana Kokoskova",slug:"dana-kokoskova"},{id:"14744",title:"Dr.",name:"Veronika",middleName:null,surname:"Venclova",fullName:"Veronika Venclova",slug:"veronika-venclova"}]},{id:"13139",title:"Herbicides in the Soil Environment: Linkage between Bioavailability and Microbial Ecology",slug:"herbicides-in-the-soil-environment-linkage-between-bioavailability-and-microbial-ecology",signatures:"M. Celina Zabaloy, Graciela P. Zanini, Virginia Bianchinotti, Marisa A. Gomez and Jay L. Garland",authors:[{id:"13386",title:"Dr.",name:"Maria Celina",middleName:null,surname:"Zabaloy",fullName:"Maria Celina Zabaloy",slug:"maria-celina-zabaloy"},{id:"14735",title:"Dr.",name:"Graciela P.",middleName:null,surname:"Zanini",fullName:"Graciela P. Zanini",slug:"graciela-p.-zanini"},{id:"14736",title:"Dr.",name:"Virginia",middleName:null,surname:"Bianchinotti",fullName:"Virginia Bianchinotti",slug:"virginia-bianchinotti"},{id:"14737",title:"Dr.",name:"Marisa",middleName:null,surname:"Gomez",fullName:"Marisa Gomez",slug:"marisa-gomez"},{id:"23759",title:"Dr.",name:"Jay",middleName:null,surname:"Garland",fullName:"Jay Garland",slug:"jay-garland"}]},{id:"13140",title:"Application of Mutated Acetolactate Synthase Genes for Herbicide Resistance to Crop Improvement",slug:"application-of-mutated-acetolactate-synthase-genes-for-herbicide-resistance-to-crop-improvement",signatures:"Masanori Shimizu, Kiyoshi Kawai, Koichiro Kaku, Tsutomu Shimizu and Hirokazu Kobayashi",authors:[{id:"14759",title:"Dr.",name:"Hirokazu",middleName:null,surname:"Kobayashi",fullName:"Hirokazu Kobayashi",slug:"hirokazu-kobayashi"}]},{id:"13141",title:"Transgenic Tall Fescue and Maize with Resistance to ALS-Inhibiting Herbicides",slug:"transgenic-tall-fescue-and-maize-with-resistance-to-als-inhibiting-herbicides",signatures:"Hiroko Sato, Tadashi Takamizo, Junko Horita, Kiyoshi Kawai, Koichiro Kaku and Tsutomu Shimizu",authors:[{id:"13817",title:"Dr.",name:"Hiroko",middleName:null,surname:"Sato",fullName:"Hiroko Sato",slug:"hiroko-sato"},{id:"13865",title:"Dr.",name:"Tadashi",middleName:null,surname:"Takamizo",fullName:"Tadashi Takamizo",slug:"tadashi-takamizo"},{id:"23766",title:"Dr.",name:"Junko",middleName:null,surname:"Horita",fullName:"Junko Horita",slug:"junko-horita"},{id:"23767",title:"Dr.",name:"Kiyoshi",middleName:null,surname:"Kawai",fullName:"Kiyoshi Kawai",slug:"kiyoshi-kawai"},{id:"23768",title:"Dr.",name:"Koichiro",middleName:null,surname:"Kaku",fullName:"Koichiro Kaku",slug:"koichiro-kaku"},{id:"23769",title:"Dr.",name:"Tsutomu",middleName:null,surname:"Shimizu",fullName:"Tsutomu Shimizu",slug:"tsutomu-shimizu"}]},{id:"13142",title:"Pollen Mediated Gene Flow in GM Crops: the Use of Herbicides as Markers for Detection. the Case of Wheat",slug:"pollen-mediated-gene-flow-in-gm-crops-the-use-of-herbicides-as-markers-for-detection-the-case-of-whe",signatures:"Iñigo Loureiro, Concepción Escorial, Inés Santín and Cristina Chueca",authors:[{id:"13724",title:"Dr.",name:"Maria-Cristina",middleName:null,surname:"Chueca",fullName:"Maria-Cristina Chueca",slug:"maria-cristina-chueca"},{id:"13725",title:"Dr.",name:"Ines",middleName:null,surname:"Santin-Montanya",fullName:"Ines Santin-Montanya",slug:"ines-santin-montanya"},{id:"13726",title:"Dr.",name:"Iñigo",middleName:null,surname:"Loureiro",fullName:"Iñigo Loureiro",slug:"inigo-loureiro"},{id:"13727",title:"Dr.",name:"Maria-Concepcion",middleName:null,surname:"Escorial",fullName:"Maria-Concepcion Escorial",slug:"maria-concepcion-escorial"}]},{id:"13143",title:"Overview of Analytical Techniques for Herbicides in Foods",slug:"overview-of-analytical-techniques-for-herbicides-in-foods",signatures:"Hua Kuang, Libing Wang and Chuanlai Xu",authors:[{id:"14502",title:"Prof.",name:"Chuanlai",middleName:null,surname:"Xu",fullName:"Chuanlai Xu",slug:"chuanlai-xu"},{id:"14802",title:"Dr.",name:"Hua",middleName:null,surname:"Kuang",fullName:"Hua Kuang",slug:"hua-kuang"}]},{id:"13144",title:"Enantioseparation and Enantioselective Analysis of Chiral Herbicides",slug:"enantioseparation-and-enantioselective-analysis-of-chiral-herbicides",signatures:"Lixia Jin, Weiliang Gao, Ling Li, Jing Ye, Chunmian Lin and Weiping Liu",authors:[{id:"15144",title:"Dr.",name:"Chunmian",middleName:null,surname:"Lin",fullName:"Chunmian Lin",slug:"chunmian-lin"}]},{id:"13145",title:"Residual Herbicide Dissipation in Vegetable Production",slug:"residual-herbicide-dissipation-in-vegetable-production",signatures:"Timothy Grey and William Vencill",authors:[{id:"13361",title:"Dr.",name:"William",middleName:null,surname:"Vencill",fullName:"William Vencill",slug:"william-vencill"},{id:"13772",title:"Dr.",name:"Timothy",middleName:null,surname:"Grey",fullName:"Timothy Grey",slug:"timothy-grey"}]},{id:"13146",title:"Solid-Phase Extraction for Enrichment and Separation of Herbicides",slug:"solid-phase-extraction-for-enrichment-and-separation-of-herbicides",signatures:"Pyrzynska Krystyna",authors:[{id:"13414",title:"Prof.",name:"Krystyna",middleName:null,surname:"Pyrzynska",fullName:"Krystyna Pyrzynska",slug:"krystyna-pyrzynska"}]},{id:"13147",title:"Chemometric Strategies for the Extraction and Analysis Optimization of Herbicide Residues in Soil Samples",slug:"chemometric-strategies-for-the-extraction-and-analysis-optimization-of-herbicide-residues-in-soil-sa",signatures:"Cristina Díez, Enrique Barrado and José Antonio Rodríguez",authors:[{id:"13645",title:"Dr.",name:"Cristina",middleName:null,surname:"Diez",fullName:"Cristina Diez",slug:"cristina-diez"},{id:"15104",title:"Dr.",name:"Enrique",middleName:null,surname:"Barrado",fullName:"Enrique Barrado",slug:"enrique-barrado"}]},{id:"13148",title:"Membrane Treatment of Potable Water for Pesticides Removal",slug:"membrane-treatment-of-potable-water-for-pesticides-removal",signatures:"Anastasios Karabelas and Konstantinos Plakas",authors:[{id:"14176",title:"Dr.",name:"Anastasios",middleName:null,surname:"Karabelas",fullName:"Anastasios Karabelas",slug:"anastasios-karabelas"},{id:"15404",title:"Dr.",name:"Konstantinos",middleName:"Vasileios",surname:"Plakas",fullName:"Konstantinos Plakas",slug:"konstantinos-plakas"}]},{id:"13149",title:"Eletrochemical Oxidation of Herbicides",slug:"eletrochemical-oxidation-of-herbicides",signatures:"Sidney Aquino Neto and Adalgisa Rodrigues De Andrade",authors:[{id:"13473",title:"Prof.",name:"Adalgisa",middleName:"Rodrigues",surname:"De Andrade",fullName:"Adalgisa De Andrade",slug:"adalgisa-de-andrade"},{id:"13476",title:"Prof.",name:"Sidney",middleName:null,surname:"Aquino Neto",fullName:"Sidney Aquino Neto",slug:"sidney-aquino-neto"}]},{id:"13150",title:"The Bioassay Technique in the Study of the Herbicide Effects",slug:"the-bioassay-technique-in-the-study-of-the-herbicide-effects",signatures:"Pilar Sandín-España, Iñigo Loureiro, Concepción Escorial, Cristina Chueca and Inés Santín-Montanya",authors:[{id:"13724",title:"Dr.",name:"Maria-Cristina",middleName:null,surname:"Chueca",fullName:"Maria-Cristina Chueca",slug:"maria-cristina-chueca"},{id:"13725",title:"Dr.",name:"Ines",middleName:null,surname:"Santin-Montanya",fullName:"Ines Santin-Montanya",slug:"ines-santin-montanya"},{id:"13726",title:"Dr.",name:"Iñigo",middleName:null,surname:"Loureiro",fullName:"Iñigo Loureiro",slug:"inigo-loureiro"},{id:"13727",title:"Dr.",name:"Maria-Concepcion",middleName:null,surname:"Escorial",fullName:"Maria-Concepcion Escorial",slug:"maria-concepcion-escorial"},{id:"23961",title:"Pilar",name:"Sandin",middleName:null,surname:"España",fullName:"Sandin España",slug:"sandin-espana"}]},{id:"13151",title:"Plasmodesmata: Symplastic Transport of Herbicides Within the Plant",slug:"plasmodesmata-symplastic-transport-of-herbicides-within-the-plant",signatures:"Germani Concenco and Leandro Galon",authors:[{id:"13555",title:"Dr.",name:"Germani",middleName:null,surname:"Concenco",fullName:"Germani Concenco",slug:"germani-concenco"},{id:"15476",title:"Prof.",name:"Leandro",middleName:null,surname:"Galon",fullName:"Leandro Galon",slug:"leandro-galon"}]},{id:"13152",title:"7-Keto-8-Aminopelagonic Acid Synthase as a Potential Herbicide Target",slug:"7-keto-8-aminopelagonic-acid-synthase-as-a-potential-herbicide-target",signatures:"In-Taek Hwang, Dong-Hee Lee and No-Joong Park",authors:[{id:"14070",title:"Dr.",name:"In-Taek",middleName:null,surname:"Hwang",fullName:"In-Taek Hwang",slug:"in-taek-hwang"}]},{id:"13153",title:"Possibilities of Applying Soil Herbicides in Fruit Nurseries – Phytotoxicity and Selectivity",slug:"possibilities-of-applying-soil-herbicides-in-fruit-nurseries-phytotoxicity-and-selectivity",signatures:"Zarya Rankova",authors:[{id:"13412",title:"Dr.",name:"Zarya",middleName:null,surname:"Rankova",fullName:"Zarya Rankova",slug:"zarya-rankova"}]},{id:"13217",title:"Herbicide Sulcotrione",slug:"herbicide-sulcotrione",signatures:"Nanxiang Wu",authors:[{id:"14277",title:"Dr.",name:"Nanxiang",middleName:null,surname:"Wu",fullName:"Nanxiang Wu",slug:"nanxiang-wu"},{id:"14804",title:"Senior Experimentalist",name:"Feng",middleName:null,surname:"Jin",fullName:"Feng Jin",slug:"feng-jin"},{id:"14805",title:"assistant researchers",name:"Yong",middleName:null,surname:"Jin",fullName:"Yong Jin",slug:"yong-jin"}]},{id:"13154",title:"The Hemodynamic Effects of the Formulation of Glyphosate-Surfactant Herbicides",slug:"the-hemodynamic-effects-of-the-formulation-of-glyphosate-surfactant-herbicides",signatures:"Hsin-Ling Lee and How-Ran Guo",authors:[{id:"14877",title:"Dr.",name:"How-Ran",middleName:null,surname:"Guo",fullName:"How-Ran Guo",slug:"how-ran-guo"},{id:"14881",title:"Dr.",name:"Hsin-Ling",middleName:null,surname:"Lee",fullName:"Hsin-Ling Lee",slug:"hsin-ling-lee"}]},{id:"13155",title:"Herbicides and Protozoan Parasite Growth Control: Implications for New Drug Development",slug:"herbicides-and-protozoan-parasite-growth-control-implications-for-new-drug-development",signatures:"Ricardo B. Leite, Ricardo Afonso and M. Leonor Cancela",authors:[{id:"14471",title:"Dr.",name:"M. Leonor",middleName:null,surname:"Cancela",fullName:"M. Leonor Cancela",slug:"m.-leonor-cancela"},{id:"14472",title:"MSc.",name:"Ricardo",middleName:null,surname:"B. Leite",fullName:"Ricardo B. Leite",slug:"ricardo-b.-leite"},{id:"14473",title:"Msc",name:"Ricardo",middleName:null,surname:"Afonso",fullName:"Ricardo Afonso",slug:"ricardo-afonso"}]},{id:"13156",title:"Synthesis and Evaluation of Pyrazine Derivatives with Herbicidal Activity",slug:"synthesis-and-evaluation-of-pyrazine-derivatives-with-herbicidal-activity",signatures:"Martin Doležal and Katarína Kráľova",authors:[{id:"13650",title:"Prof.",name:"Martin",middleName:null,surname:"Dolezal",fullName:"Martin Dolezal",slug:"martin-dolezal"},{id:"14876",title:"Dr.",name:"Katarina",middleName:null,surname:"Kralova",fullName:"Katarina Kralova",slug:"katarina-kralova"}]}]}]},onlineFirst:{chapter:{type:"chapter",id:"64006",title:"Cumulative Tensile Damage and Consolidation Effects on Fracture Properties of Sandstone",doi:"10.5772/intechopen.81434",slug:"cumulative-tensile-damage-and-consolidation-effects-on-fracture-properties-of-sandstone",body:'\n
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1. Introduction
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Visible cracks are present on most historical buildings, structures, and objects of art made of porous brittle or quasi-brittle materials. Stone monuments form an important category of these structures.
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In principle, visible cracks may develop due to external forces acting under various time situations, for example, incidental shocks or gradually increasing loads, or due to internal strain gradients, which mostly generate cumulative damage, such as fatigue phenomena.
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Cumulative tensile damage may occur in stone monuments due to repeated environmental uneven volumetric change. Stone objects are frequently subjected to repeated changes in temperature from solar irradiation, when the heat is transferred unevenly inside the material. The accompanying different thermal dilations may cause dangerous stresses that lead to cumulative crack propagation, typically initiated in the interior defects that are usually present. In addition, a combination of temperature and moisture dilation effects may worsen the situation, especially in sandstone or in specific fragile stones, e.g., in the so-called “opuka” stone (Cretaceous marly stone) used in the Czech Republic. In the case of these materials, stone elements in the interior environment are also threatened by environmental fatigue. Let us present a typical example that has been described in greater detail in Drdácký [1].
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The second author of this paper investigated the case of a severe failure of the sandstone tracery of the triforium of St. Vitus Cathedral, where repeated uneven humidity loading (wetting and drying) and temperature loading (heating and cooling) were the cause of the collapse. A detailed survey discovered that this type of damage was present or had been repaired on most of the tracery arches of the triforium, where further failures subsequently occurred. The extent of the dangerous stresses was verified using simple thermal loading modeling and computations. It was proved that the state of stress in the damaged places reached or surpassed the material strength levels (Figure 1) [2].
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Figure 1.
Distribution of the effective stress in the slice for coordinate y = 1.37; 3D model of own weight (left) and 3D model of warming up (right) [1, 2]. (Image by P. Beran).
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The triforium tracery elements are chiseled out of a single piece of sandstone. They are connected with the lower window beam of the nave or the transept. The triforium structure is situated on the border between the exterior and the interior, and its ceiling plates are exposed to the exterior environment forming the roof structure. The temperature of the air fluctuates in the exterior and the interior of the cathedral. The stone elements of the triforium include parts of different proportions that have been composed into a single unit. Due to rapid changes in the ambient temperature, situations can arise when the temperature of the massive parts is different from the temperature of the subtler elements. This situation generates a stress inside the structure. The same effect is caused by the change in moisture content that always accompanies temperature changes.
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Similar damage has been observed on other historic buildings, for example, on the church of Notre Dame du Sablon in Brussels (see Figure 2) where several failed original elements have been replaced.
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Figure 2.
Repaired tracery with damaged elements in Notre Dame du Sablon (photo by M. Drdácký).
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Adverse effects of this type have led to a call for studies of the behavior of similar stone structures under extreme climate effects. These studies require greater knowledge than what is available of the fracture characteristics of the historical stone. Numerous works on the fracture behavior of rock have been published, mostly based on tests on cylindrical stone specimens under cyclic compression according to the ISRM standards (e.g., Ning et al. [3], Bagde and Petroš [4], Backers [5]). Quantitative toughness data are calculated, or qualitative data are assessed on the basis of these tests. A classical work by Attewell and Farmer [6] and also a comprehensive work by Jian-Qing et al. [7], which review fatigue damage variables and discuss a broad range of NDT measurement methods, in particular ultrasonic and acoustic emission methods, are worth mentioning. However, this type of testing methodology is not suitable for studies of the effects of conservation treatment on the fatigue behavior of stone. No pure tensile fatigue tests on stone have been found in the literature, only three-point or four-point bending tests (e.g., Cardani and Meda [8]). There is a lack of data on the behavior of stone under repeated uniaxial tensile loads, and until now no suitable methodology for testing stone toughness before and after conservation treatment has been suggested and accepted.
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This paper presents the results of an experimental pilot assessment of changes in the mechanical properties of sandstone from Božanov (a typical material used on the Charles Bridge in medieval times) resulting from accumulated damage. The Young modulus and the Poisson number were observed. In addition, a methodology for testing stone in tension and in cyclic tension/compression loading is suggested and verified.
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2. Experiments
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Božanov stone is a grayish beige gross grain strong arkose sandstone without marked layering (Figure 3). The material used in the tests was extracted from the 11th arch of the Charles Bridge parapet wall, which had to be replaced during recent repairs. It has a rather deeply located detachment crack parallel to the surface, probably due to some previous surface treatment which had locked moisture inside the stone. The material can be characterized as a quasi-brittle inelastic silicate composite.
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Figure 3.
Macro photo of the Božanov sandstone (arkose) structure (photo J. Frankl).
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Test specimens 50 × 50 × 200 mm3 in dimensions were fixed into rectangular steel tubes with axially welded flat steel hangers, which served to fix the set into the hydraulic grips of the loading frame (Figure 4). Two-component Sikadur-31 CF RAP resin was used for gluing. The fixtures had to be prepared with great precision in order to ensure perfect alignment and perpendicular arrangement for tension and combined loading. The specimens in prismatic form were cut with negligible geometrical imperfections in a precise prismatic form.
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Figure 4.
Test compression/tension specimens (photo by M. Šperl).
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The specimens were loaded into an Instron 1343 electrohydraulic testing frame with force capacity of 100 kN (Figure 5) using an Instron 100 kN load cell, a FastTrack 8800 controller, two Instron extensometers type 2620-602 (accuracy class 1, with maximum possible error of 0.5% of the read value, measurement base l0 = 50 mm), and WaveMatrix measurement software. The extensometers were placed on opposite sides of the specimen, and their read values were arithmetically averaged. The contacts on the surface of the stone were provided with glued thin aluminum sheets to protect the extensometer wedges. The deformations were also recorded and evaluated using a contactless telecentric digital camera and the digital image correlation (DIC) method. This method utilizes a sequence of acquired images that represents a specimen surface deformation process. In this sequence, DIC observes the displacements of small rectangular regions (templates) on the sample with a distinguishable distribution of gray-scale intensities [9, 10]. The displacements obtained from this method are utilized for calculating the strains. In the case of stone, the random surface pattern, a prerequisite for the method, is formed by the distinct natural texture of the surface of the specimen.
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Figure 5.
Arrangement of pure tension tests on the stone specimens (photo by M. Šperl).
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The DIC algorithm used in this work has two main steps. First, an integer value of the pixel displacement is evaluated using a normalized cross-correlation function for a central pixel of a square image template in the reference image. The second step in the matching process is known as the Lucas-Kanade algorithm [11]. This step takes into account the reference template’s own deformation. The method searches for an affine transformation that projects the template onto the deformed image using minimization of the sum-squared differences between the template and the deformed image. The LK algorithm is an iterative nonlinear optimization method. The integer values of the pixel displacement estimated in the first step were passed to this iterative process as initial guess values. The longitudinal deformation was measured using two rows of points. Each point represents the center of the rectangular template, the displacement of which was measured by DIC. Multiple points were used, and their displacements were finally averaged for noise reduction. Subsequently, the longitudinal engineering strain was calculated using the measured displacements as
\n\n\nE1
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Knowing the measured applied force and the dimensions of the cross section of the specimen, the engineering stress can finally be evaluated.
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Twelve experiments were carried out—four on the specimens from Figure 4 and eight on prismatic beams cut out of the broken parts after the tension tests. The tests therefore involved (i) compression, (ii) very-low-cycle fatigue, (iii) tension, (iv) fatigue (an alternating nonsymmetrical cycle)—all on the specimens from Figure 4—and (v) three-point bending tests on small size beams with a 20 × 20 mm2 cross section.
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The effect of consolidation on the fracture behavior was studied on prismatic beams of the same cross section as described above provided in its center with a notch and a generated crack. The crack generation procedure is described in detail by Drdácký et al. [12]. Specimens with similar length of crack were consolidated applying the most typical agents: elastified Steinfestiger 300 (30% concentration of the active substance), Paraloid B-72 (2% concentration), and Funcosil 100 (10% concentration of the active substance). After maturing they were tested in standard static three-point bending.
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3. Test results
\n
The compression force was applied in three stages: −1.5, −26, and −37 MPa. Deformation was measured during the tests in order to study the change in Young’s modulus and the Poisson number with relation to applied load (Figures 6 and 7). Hysteresis and irreversible deformation were observed, and the Poisson number reached a value of 0.23. The diagram shows that the tested sandstone increased in stiffness with an increasing number of compression cycles at the beginning of the tests. The change in the slope of the interpolated linear approximations of the load-displacement diagrams reflects this fact. The modulus of elasticity rose from an initial value of 11,360 MPa to a value of 19,237 MPa for the third cycle of loading, i.e., a 70% increase. This differs from the results of other tests on sandstone as presented in the cited literature. At the same time, hysteresis occurs with irreversible deformations. During the first loading cycle to −1.5 MPa, no irreversible deformation was observed. It seems that irreversible deformation occurs after the material has been subjected to some limit compression load. The Czech standard for tests on natural stone prescribes that the modulus of elasticity is to be evaluated after three loading cycles.
\n
Figure 6.
First compression loading cycle (left) and second and third loading cycles (right), with corresponding linear approximations.
\n
Figure 7.
Poisson ratio measured using the DIC method.
\n
During the tension tests, the velocity of the crosshead movement was intentionally kept very low at a value of 5 μm/min. Due to this low loading velocity, it was possible to evaluate the overall energy absorbed during crack origin and propagation upon the initiation of a crack at a defect present within the specimen (Figure 8). The load-displacement graph shows the overall energy absorbed during the test (0.4 J) and during crack growth (0.16 J). This corresponds to the behavior of highly brittle material. The strength decreased (to 1.43 MPa) due to the defect in the material. The course of the stress and deformation up to fracture can be described well by means of a suitable power function (see Figure 9), which presents a detailed diagram.
\n
Figure 8.
Load-crosshead displacement diagram for the tension test with a calculation of the absorbed energies and an indication of the probable place with an initial defect in the cross section of the specimen.
\n
Figure 9.
Tension diagram of the tested Božanov sandstone, comparing the conservative measurement with the DIC measurement and presenting a suitable power function approximation.
\n
It follows from Figure 9 that a suitable power function appropriately describes the material behavior up to rupture. The tangent to the curve gives the modulus of elasticity value:
\n\n\nE2
\n
The power function seems to be ideal for describing the deformation behavior of this sandstone.
\n
The DIC record exhibits a slight difference in comparison with the conservative measurement approach. This may be caused by a small change in the geometric relations between the specimen and the camera objective during loading. However, the agreement is still very good.
\n
A sinusoidal altering nonsymmetrical loading cycle with stress limits of +1.5 and −2MPa was applied during the very-low-fatigue test. The mean value of the stress was −0.25 MPa, the stress double amplitude was 3.5 MPa, and the frequency of loading was 0.2 Hz (Figure 10).
\n
Figure 10.
Low-cycle fatigue diagram.
\n
The maximum tension stress (σmax ≈ 1.5 MPa) approached the limit stress (strength) in tension of the tested material (the static strength was 1.43 MPa on a weakened profile; see above). The resulting number of cycles was only 1.25; the fracture occurred while the maximum tension load was being attained in the second cycle.
\n
Figure 10 also indicates that a power function can be used to approximate the first loading course. The parameters of the constant however are different from that of the previous case. This could be a result of the presence of an interior defect in the first case or merely the wide distribution of the material’s characteristics. The change in the fatigue loop’s tension branch is also noteworthy. In comparison to the first cycle, it is significantly straighter and more horizontal. It is even possible to approximate it with a linear function. It signifies considerable damage to the material resulting from the first loading cycle, corresponding to the stress applied. The hysteresis exhibited by this specimen is quite high. The compression component is steeper than the tension component [14].
\n
The nonsymmetrical altering sinusoidal loading cycle was also used in the fatigue tests, with stress limits of +0.265 and −0.354 MPa. There were two modes of cycling velocity used—for the main loading sequence a frequency of 0.25 and 0.01 Hz at certain stages for groups of three cycles measuring deformations, based on which the second cycle was evaluated and the E modulus calculated. Such detailed measurements were attained up to 2000 cycles. The specimen was then loaded to the point of failure. The asymmetry characteristics of the cycle were kept the same to allow for comparability of results (R = −1.33 and stress span Δσ = 0.619 MPa). The overall lifespan reached 224,908 loading cycles. Typical results are presented in Figure 11.
\n
Figure 11.
Stress-strain ratios after a given number of cycles: 22 (top left), 42 (top right), 82 (center left), 202 (center right), 602 (bottom left), and 2002 (bottom right).
\n
The modulus of elasticity E changed during the cycling. Its value dropped from the initial figure of 11,214–10,705 MPa after 82 loading cycles (a decrease of about 5%). Then the value stabilized around 10,865 MPa after 2002 cycles (Figure 12). The change was influenced by fatigue damage in the material.
\n
Figure 12.
Change in the modulus of elasticity during cycling.
\n
In Figure 11, the observed shift of the measured data toward negative strain values with the increasing number of cycles is caused by thermal dilation of the specimens. The average change in temperature in the testing hall was about 4°C. The temperature change dilations were checked by computations, taking into account the coefficient of dilation of sandstone equal to α = 10 × 10−6 K−1 and the equation Δl = α·ΔT·l. Figure 13 presents examples of the temperature drift.
\n
Figure 13.
Temperature caused a drift in specimen deformation due to dilation (2000–20,000 cycles).
\n
Figure 14 presents a very rough representation of the probable life estimate of the tested sandstone between two applied stress levels. Of course, many more tests will be required before the true Wöhler curve can be constructed.
\n
Figure 14.
Fatigue test results for Božanov sandstone for cycle asymmetry of R = −1.33.
\n
After the uniaxial and fatigue tests, small beams with a cross section of 20 × 20 mm2 were prepared from the broken specimens and were subjected to three-point bending loading (Figure 15). The measured flexural strength reached much higher values (about 4.5 MPa) than the strength in the tension tests, which was influenced by a rather low span to height ratio (about 3.75).
\n
Figure 15.
Three-point bending test arrangement (photo O. Vála).
\n
However, the moduli of elasticity were lower than those measured during the previous tests (though still higher than those measured by the first author on thin plates during tests on the characteristics of the Charles Bridge sandstone—Čechová et al. [13]). It is clear that both the load-displacement diagram and the change in the modulus of elasticity can be well approximated by a suitable mathematical model (Figure 16).
\n
Figure 16.
Loading force-deflection diagram during three-point bending testing and calculated modulus of elasticity.
\n
In the study on consolidation effects influencing crack propagation and toughness, three types of specimens were tested—notched beams without cracks, notch beams with cracks generated by cycling, and notched beams with cracks after consolidation [14]. They were loaded in three-point bending configuration.
\n
The results show that consolidation increased the load-carrying capacity of the cracked sandstone specimens. The reference specimen with the initiation notch attained an average bending strength of 5.05 MPa, the notched specimen with the cycled crack one of 4.56 MPa, and the consolidated specimen one of about 6 MPa.
\n
\n
\n
4. Conclusions
\n
Although it is difficult to generalize the results of the pilot tests described here, they provide important findings for further planning of stone fatigue tests.
\n
The methodology of tension tests using specimens inserted and glued in tubular fixtures is functional and has been verified. However, it is very demanding from the point of view of specimen preparation, and it requires very skilled staff. It can be recommended mainly for special small-series tests. For research requiring tests on a large numbers of specimens, it is necessary to develop some other techniques. However, the pilot tests were successfully completed, and the energies for crack initiation and propagation in the given sandstone have been determined.
\n
The optical DIC method again proved its capacity and its suitability for measurements of a complex deformation field on porous and naturally well-structured surfaces.
\n
The modulus of elasticity may be a good parameter for assessing damage accumulation, though it is difficult to measure on real structures (see Figure 11). In the next series of tests, the correlation between the modulus of elasticity and NDT techniques will be studied, especially ultrasonic measurements for possible application in situ.
\n
It follows from Figure 13 that the first tension load displacement can be approximated very satisfactorily by means of a power function.
\n
The tests with consolidated specimens proved the positive effects of stone impregnation on strength and toughness. All notched and cracked specimens after consolidation with ethyl silicates KSE 100 and KSE 300 and Paraloid B-72 attained values above full profile strength which was tested on identical beams. The KSE 300 consolidation agent attained the best result in the range of 145% as far as strength is concerned and 161% in the toughness value (KIC).
\n
The toughness of untreated notched and cracked specimens reached almost the same values (0.33:0.35 MPa√m), which mean that with such a heterogeneous material, one cannot expect stress concentration factors similar to those of metals.
\n
\n
Acknowledgments
\n
The authors acknowledge the kind support of the Czech Science Foundation Project GAČR P105/12/G059 and the kind collaboration with Dr. Ivan Jandejsek who made the DIC measurements and evaluated the data.
\n
\n',keywords:"damage cumulation, sandstone, historical monuments, simple tension, cyclic loading, stone consolidation",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/64006.pdf",chapterXML:"https://mts.intechopen.com/source/xml/64006.xml",downloadPdfUrl:"/chapter/pdf-download/64006",previewPdfUrl:"/chapter/pdf-preview/64006",totalDownloads:221,totalViews:97,totalCrossrefCites:0,dateSubmitted:"June 4th 2018",dateReviewed:"September 10th 2018",datePrePublished:"November 5th 2018",datePublished:"March 13th 2019",readingETA:"0",abstract:"The presence of cracks in many historical objects indicates the action of external forces accompanied by internal strain gradients. This is usually a repetitive process, and damage cumulation may occur. A study of these effects requires a suitable methodology for testing historical stone that has been subjected to repeated tension strains. The chapter presents the results of a pilot experimental assessment of changes in the mechanical characteristics of sandstone due to accumulation of damage. The Young modulus and the Poisson number were investigated, using a verified methodology for testing stone in simple tension and in cyclic simple tension/compression loading. The results show that the first tension load displacement can be approximated very satisfactorily by a power function, and the optical digital image correlation (DIC) method again demonstrated its capacity and suitability for measuring the complex deformation field on porous surfaces and on naturally well-structured surfaces. The chapter further presents a methodology for investigating fracture phenomena in sandstone treated for consolidation. It shows the preparation of test specimens with a cyclic loading generated crack, control of the test specimen preparation, and verification by means of X-ray micro-CT and DIC techniques. The chapter illustrates an influence of various consolidation agents on the toughness of cracked specimens.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/64006",risUrl:"/chapter/ris/64006",signatures:"Martin Šperl and Miloš Drdácký",book:{id:"8412",title:"Sustainable Construction and Building Materials",subtitle:null,fullTitle:"Sustainable Construction and Building Materials",slug:"sustainable-construction-and-building-materials",publishedDate:"March 13th 2019",bookSignature:"Sayed Hemeda",coverURL:"https://cdn.intechopen.com/books/images_new/8412.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"258282",title:"Prof.",name:"Sayed",middleName:null,surname:"Hemeda",slug:"sayed-hemeda",fullName:"Sayed Hemeda"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:[{id:"144548",title:"Dr.",name:"Martin",middleName:null,surname:"Šperl",fullName:"Martin Šperl",slug:"martin-sperl",email:"sperl@itam.cas.cz",position:null,institution:null},{id:"261550",title:"Prof.",name:"Miloš",middleName:null,surname:"Drdácký",fullName:"Miloš Drdácký",slug:"milos-drdacky",email:"drdacky@itam.cas.cz",position:null,institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Experiments",level:"1"},{id:"sec_3",title:"3. Test results",level:"1"},{id:"sec_4",title:"4. Conclusions",level:"1"},{id:"sec_5",title:"Acknowledgments",level:"1"}],chapterReferences:[{id:"B1",body:'Drdácký MF. Impact of climate change on building structures. In: Lefèvre RA, Sabbioni C, editors. Climate Change and Cultural Heritage. Scienze e materiali del patrimonio culturale, 10. Bari: EDIPUGLIA, s. r. l.; 2010. pp. 139-153\n'},{id:"B2",body:'Beran P, Drdácký M. Influence of temperature changes on stresses in the Triforium tracery of St Vitus’ Cathedral in Prague. In: Oñate E, Papadrakakis M, Schrefler B, editors. Proceedings Computational Methods for Coupled Problems in Science and Engineering II Coupled Problems. Barcelona: CIMNE; 2007. pp. 433-436\n'},{id:"B3",body:'Ning L, Ping Z, Yunsheng C, Gunter S. Fatigue properties of cracked, saturated and frozen sandstone samples under cyclic loading. International Journal of Rock Mechanics and Mining Sciences. 2003;40:145-150\n'},{id:"B4",body:'Bagde MN, Petroš MN. Fatigue properties of intact sandstone samples subjected to dynamic uniaxial cyclical loading. International Journal of Rock Mechanics and Mining Sciences. 2005;42:237-250\n'},{id:"B5",body:'Backers T. Fracture Toughness Determination and Micromechanics of Rock Under Mode I and Mode II Loading. PhD Thesis. Universität Potsdam 2004. Scientific Technical Report STR 05/05, GeoForschungsZentrum Potsdam; 2004. p. 138\n'},{id:"B6",body:'Attewell PB, Farmer IW. Fatigue behaviour of rock. International Journal of Rock Mechanics and Mining Sciences and Geomechanics Abstracts. 1973;10(1):1-9\n'},{id:"B7",body:'Jian-Qing X, De-Xin D, Fu-Liang J, Gen X. Fatigue damage variable and evolution of rock subjected to cyclic loading. International Journal of Rock Mechanics and Mining Science. 2010;47:461-468. DOI: 10.1016/j.ijrmms.2009.11.003\n'},{id:"B8",body:'Cardani G, Meda A. Marble behaviour under monotonic and cyclic loading in tension. Construction and Building Materials. 2004;18:419-424\n'},{id:"B9",body:'Lin Q, Labuz JF. Fracture of sandstone characterized by digital image correlation. International Journal of Rock Mechanics and Mining Sciences. 2013;60:235-245\n'},{id:"B10",body:'Peters WH, Ranson WF. Digital imaging techniques in experimental stress analysis. Optical Engineering. 1982;21:427-431\n'},{id:"B11",body:'Lucas BD, Kanade T. An iterative image registration technique with an application to stereo vision. In: Proceedings of Imaging Understanding Workshop; 1981. pp. 121-130\n'},{id:"B12",body:'Drdácký M, Šperl M, Jandejsek I. Consolidation effects on sandstone toughness. In: Hughes J, Howind T, editors. Proceedings of the 13th Int. Congress on the Deterioration and Conservation of Stone “SCIENCE and ART: A Future for Stone”. Vol. 2. Paisley: University of West Scotland; 2016. pp. 687-694\n'},{id:"B13",body:'Čechová E, Drdácký M, Frankeová D, Lesák J, Slížková Z, Valach J, et al. Technology Guidelines for Restoration of the XIth Arch of the Charles Bridge in Prague. Research Report of ITAM AS CR. Prague: ITAM; 2010. p. 139\n'},{id:"B14",body:'Šperl M, Drdácký M. Non-standard experimental tests of sandstone and its pre-cracking for fracture testing. In: IOP Conference Series: Materials Science and Engineering 379; 2018. DOI: 10.1088/1757-899X/379/1/012029\n'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"Martin Šperl",address:"sperl@itam.cas.cz",affiliation:'
Institute of Theoretical and Applied Mechanics of the Academy of Sciences of the Czech Republic, Prague, The Czech Republic
Institute of Theoretical and Applied Mechanics of the Academy of Sciences of the Czech Republic, Prague, The Czech Republic
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IntechOpen is a dynamic, vibrant company, where exceptional people are achieving great things. We offer a creative, dedicated, committed, and passionate environment but never lose sight of the fact that science and discovery is exciting and rewarding. We constantly strive to ensure that members of our community can work, travel, meet world-renowned researchers and grow their own career and develop their own experiences.
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Integrity - We are consistent and dependable, always striving for precision and accuracy in the true spirit of science.
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Openness - We communicate honestly and transparently. We are open to constructive criticism and committed to learning from it.
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Disruptiveness - We are eager for discovery, for new ideas and for progression. We approach our work with creativity and determination, with a clear vision that drives us forward. We look beyond today and strive for a better tomorrow.
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What makes IntechOpen a great place to work?
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IntechOpen is a dynamic, vibrant company, where exceptional people are achieving great things. We offer a creative, dedicated, committed, and passionate environment but never lose sight of the fact that science and discovery is exciting and rewarding. We constantly strive to ensure that members of our community can work, travel, meet world-renowned researchers and grow their own career and develop their own experiences.
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If this sounds like a place that you would like to work, whether you are at the beginning of your career or are an experienced professional, we invite you to drop us a line and tell us why you could be the right person for IntechOpen.
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